Methods for the treatment of metabolic disorders such as diabetes

ABSTRACT

Compounds useful for the treatment of various metabolic disorders, such as insulin resistance syndrome, diabetes, hyperlipidemia, fatty liver disease, cachexia, obesity, atherosclerosis and arteriosclerosis, are disclosed.

CROSS-REFERENCE TO PRIOR APPLICATIONS

This is a divisional of U.S. application Ser. No. 10/481,042, filed Jan.14, 2005, which is the U.S. national phase under 35 U.S.C. §371 ofInternational Application No. PCT/US02/18388, filed Jun. 12, 2002, thecontents of which are incorporated herein by reference. This applicationclaims the benefit of U.S. Provisional Patent Application No.60/297,282, filed Jun. 12, 2001, the content of which is incorporatedherein by reference.

BACKGROUND OF THE INVENTION

Diabetes mellitus is a major cause of morbidity and mortality.Chronically elevated blood glucose leads to debilitating complications:nephropathy, often necessitating dialysis or renal transplant;peripheral neuropathy; retinopathy leading to blindness; ulceration ofthe legs and feet, leading to amputation; fatty liver disease, sometimesprogressing to cirrhosis; and vulnerability to coronary artery diseaseand myocardial infarction.

There are two primary types of diabetes. Type I, or insulin-dependentdiabetes mellitus (IDDM) is due to autoimmune destruction ofinsulin-producing beta cells in the pancreatic islets. The onset of thisdisease is usually in childhood or adolescence. Treatment consistsprimarily of multiple daily injections of insulin, combined withfrequent testing of blood glucose levels to guide adjustment of insulindoses, because excess insulin can cause hypoglycemia and consequentimpairment of brain and other functions.

Type II, or noninsulin-dependent diabetes mellitus (NIDDM) typicallydevelops in adulthood. NIDDM is associated with resistance ofglucose-utilizing tissues like adipose tissue, muscle, and liver, to theactions of insulin. Initially, the pancreatic islet beta cellscompensate by secreting excess insulin. Eventual islet failure resultsin decompensation and chronic hyperglycemia. Conversely, moderate isletinsufficiency can precede or coincide with peripheral insulinresistance. There are several classes of drugs that are useful fortreatment of NIDDM: 1) insulin releasers, which directly stimulateinsulin release, carrying the risk of hypoglycemia; 2) prandial insulinreleasers, which potentiate glucose-induced insulin secretion, and mustbe taken before each meal; 3) biguanides, including metformin, whichattenuate hepatic gluconeogenesis (which is paradoxically elevated indiabetes); 4) insulin sensitizers, for example the thiazolidinedionederivatives rosiglitazone and pioglitazone, which improve peripheralresponsiveness to insulin, but which have side effects like weight gain,edema, and occasional liver toxicity; 5) insulin injections, which areoften necessary in the later stages of NIDDM when the islets have failedunder chronic hyperstimulation.

Insulin resistance can also occur without marked hyperglycemia, and isgenerally associated with atherosclerosis, obesity, hyperlipidemia, andessential hypertension. This cluster of abnormalities constitutes the“metabolic syndrome” or “insulin resistance syndrome”. Insulinresistance is also associated with fatty liver, which can progress tochronic inflammation (NASH; “nonalcoholic steatohepatitis”), fibrosis,and cirrhosis. Cumulatively, insulin resistance syndromes, including butnot limited to diabetes, underlie many of the major causes of morbidityand death of people over age 40.

Despite the existence of such drugs, diabetes remains a major andgrowing public health problem. Late stage complications of diabetesconsume a large proportion of national health care resources. There is aneed for new orally active therapeutic agents which effectively addressthe primary defects of insulin resistance and islet failure with feweror milder side effects than existing drugs.

Currently there are no safe and effective treatments for fatty liverdisease. Therefore such a treatment would be of value in treating thiscondition.

SUMMARY OF THE INVENTION

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein n is 1 or 2; m is 0 or 1; q is 0 or 1; t is 0 or 1; R⁵ is alkylhaving from 1 to 3 carbon atoms; R⁹ is hydrogen, halo, or alkoxy havingfrom 1 to 3 carbon atoms;A is phenyl, unsubstituted or substituted by 1 or 2 groups selectedfrom: halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl, alkoxyhaving 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkyl havingfrom 3 to 6 ring carbon atoms wherein the cycloalkyl is unsubstituted orone or two ring carbons are independently mono-substituted by methyl orethyl; or a 5 or 6 membered heteroaromatic ring having 1 or 2 ringheteroatoms selected from N, S and O and the heteroaromatic ring iscovalently bound to the remainder of the compound of formula I′ by aring carbon; and X is—CH₂—, Q is—OR¹ and R¹ is ethyl; or X is —CH₂CR¹²R¹³— or —CH₂CH(NHAc)— wherein eachof R¹² and R¹³ is independently hydrogen or methyl, Q is OR¹ and R¹ ishydrogen or alkyl having from 1 to 7 carbon atoms; or X is —CH₂CH₂— andQ is NR¹⁰R¹¹ wherein one of R¹⁰ and R¹¹ is hydrogen, alkyl having from 1to 3 carbon atoms or hydroxy, and the other is hydrogen or alkyl havingfrom 1 to 3 carbon atoms; or when R¹ is hydrogen, a pharmaceuticallyacceptable salt of the compound.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein n is 1 or 2; t is 0 or 1; m is 0 and r is 1, or m is 1 and r is0; A is phenyl, unsubstituted or substituted by 1 or 2 groups selectedfrom: halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl, alkoxyhaving 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkyl havingfrom 3 to 6 ring carbon atoms wherein the cycloalkyl is unsubstituted orone or two ring carbons are mono-substituted by methyl or ethyl; or a 5or 6 membered heteroaromatic ring having 1 or 2 ring heteroatomsselected from N, S and O and the heteroaromatic ring is covalently boundto the remainder of the compound of formula II by a ring carbon; Z is

R¹ is hydrogen or alkyl having from 1 to 7 carbon atoms; R⁴ is hydrogen;—NHCOOC(CH₃)₃; —NHCH₃; or —NHCH₂CH₃; or when R¹ is hydrogen, apharmaceutically acceptable salt of the compound.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

whereinn is 1 or 2; A is phenyl, unsubstituted or substituted by 1 or 2 groupsselected from: halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl,alkoxy having 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkylhaving from 3 to 6 ring carbon atoms wherein one or both ring carbonsare independently mono-substituted by methyl or ethyl; or a 5 or 6membered heteroaromatic ring having 1 or 2 ring heteroatoms selectedfrom N, S and O and the heteroaromatic ring is covalently bound to theremainder of the compound of formula III by a ring carbon.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein R¹ is hydrogen or alkyl having from 1 to 7 carbon atoms; or whenR¹ is hydrogen, a pharmaceutically acceptable salt of the compound.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein n is 1 or 2; R¹ is hydrogen or alkyl having from 1 to 7 carbonatoms; R¹⁴ is hydroxy or hydrogen; and A is phenyl, unsubstituted orsubstituted by 1 or 2 groups selected from halo, alkyl having 1 or 2carbon atoms, perfluoromethyl, alkoxy having 1 or 2 carbon atoms, andperfluoromethoxy; or cycloalkyl having from 3 to 6 ring carbon atomswherein the cycloalkyl is unsubstituted or one or two ring carbons areindependently mono-substituted by methyl or ethyl; or a 5 or 6 memberedheteroaromatic ring having 1 or 2 ring heteroatoms selected from N, Sand O and the heteroaromatic ring is covalently bound to the remainderof the compound of formula V′ by a ring carbon; or a pharmaceuticallyacceptable salt of the compound.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein n is 1 or 2; R¹ is hydrogen or alkyl having from 1 to 3 carbonatoms; and A is phenyl, unsubstituted or substituted by 1 or 2 groupsselected from halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl,alkoxy having 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkylhaving from 3 to 6 ring carbon atoms wherein the cycloalkyl isunsubstituted or one or two ring carbons are independentlymono-substituted by methyl or ethyl; or a 5 or 6 membered heteroaromaticring having 1 or 2 ring heteroatoms selected from N, S and O and theheteroaromatic ring is covalently bound to the remainder of the compoundof formula XCI by a ring carbon; or a pharmaceutically acceptable saltof the compound.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein n is 1 or 2; R¹ is hydrogen or alkyl having from 1 to 3 carbonatoms; and A is phenyl, unsubstituted or substituted by 1 or 2 groupsselected from halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl,alkoxy having 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkylhaving from 3 to 6 ring carbon atoms wherein the cycloalkyl isunsubstituted or one or two ring carbons are independentlymono-substituted by methyl or ethyl; or a 5 or 6 membered heteroaromaticring having 1 or 2 ring heteroatoms selected from N, S and O and theheteroaromatic ring is covalently bound to the remainder of the compoundof formula CXVI by a ring carbon; or a pharmaceutically acceptable saltof the compound.

This invention provides a biologically active agent, wherein the agentis a compound of the formula:

wherein n is 0, 1 or 2; R¹ is hydrogen or alkyl having from 1 to 3carbon atoms; R¹⁵ is hydrogen or alkyl having from 1 to 3 carbon atoms;R⁹ is hydrogen, halo, hydroxy, or alkoxy having from 1 to 3 carbonatoms; A is phenyl, unsubstituted or substituted by 1 or 2 groupsselected from halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl,alkoxy having 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkylhaving from 3 to 6 ring carbon atoms wherein the cycloalkyl isunsubstituted or one or two ring carbons are independentlymono-substituted by methyl or ethyl; or a 5 or 6 membered heteroaromaticring having 1 or 2 ring heteroatoms selected from N, S and O and theheteroaromatic ring is covalently bound to the remainder of the compoundof formula CXVII by a ring carbon; or a pharmaceutically acceptable saltof the compound.

The biologically active agents described above have activity in one ormore of the biological activity assays described below, which areestablished animal models of human diabetes and insulin resistancesyndrome. Therefore such agents would be useful in the treatment ofdiabetes and insulin resistance syndrome. All of the exemplifiedcompounds that were tested demonstrated activity in the biologicalactivity assay or assays in which they were tested.

This invention provides the use of the biologically active agentsdescribed above in the manufacture of a medicament for the treatment ofinsulin resistance syndrome, diabetes, cachexia, hyperlipidemia, fattyliver disease, obesity, atherosclerosis or arteriosclerosis. Thisinvention also provides methods of treating a mammalian subject withinsulin resistance syndrome, diabetes, cachexia, hyperlipidemia, fattyliver disease, obesity, atherosclerosis or arteriosclerosis comprisingadministering to the subject an effective amount of a biologicallyactive agent in accordance with this invention. This invention alsoprovides a pharmaceutical composition comprising a biologically activeagent of this invention and a pharmaceutically acceptable carrier.

This invention provides certain novel intermediates, which are useful inproducing the biologically active agents of this invention. Theinvention also provides processes of producing the biologically activeagents and intermediates.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1: Serum insulin levels in high fat-fed C57Bl/6J mice receivingvehicle (negative control), Compound BI, Compound BL, Wy14643 orrosiglitazone.

FIG. 2: Serum leptin levels in high fat-fed C57Bl/6J mice receivingvehicle (negative control), Compound BI, Compound BL, Wy14643 orrosiglitazone.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

As used herein the term “alkyl” means a linear or branched-chain alkylgroup. An alkyl group identified as having a certain number of carbonatoms means any alkyl group having the specified number of carbons. Forexample, an alkyl having three carbon atoms can be propyl or isopropyl;and alkyl having four carbon atoms can be n-butyl, 1-methylpropyl,2-methylpropyl or t-butyl.

As used herein the term “halo” refers to one or more of fluoro, chloro,bromo, and iodo.

As used herein the term “perfluoro” as in perfluoromethyl orperfluoromethoxy, means that the group in question has fluorine atoms inplace of all of the hydrogen atoms.

As used herein “Ac” refers to the group CH₃C(O)—.

Examples of the biologically active compounds of the instant inventionare listed below. These compounds are referred to herein by theirchemical name or by the two-letter code shown below.

-   AJ 4-(4-(2-(2-Fluorophenyl)ethoxy)phenyl)-4-oxobutyric acid;-   AK Ethyl 4-(4-(2-fluorobenzyloxyl)phenyl)-4-oxobutyrate;-   AL 4-(4-(2-Methylbenzyloxy)phenyl)-4-oxobutyric acid;-   AM    4-[4-(2-(N-(2-fluorobenzyl)-N-methylamino)ethoxy)phenyl]-4-oxobutyric    acid;-   AN 4-(3-(2-Methylbenzyloxy)phenyl)-4-oxobutyric acid;-   AO Ethyl 4-(3-(2-fluorobenzyloxyl)phenyl)-4-oxobutyrate;-   AP Ethyl 4-(4-(2-methylbenzyloxyl)phenyl)-4-oxobutyrate;-   AQ Ethyl 4-(4-(2,6-difluorobenzyloxyl)phenyl)-4-oxobutyrate;-   AR 4-(4-(2-(2-Thienyl)ethoxy)phenyl)-4-oxobutyric acid;-   AS 4-(2,6-Difluorophenyl)-4-oxobutyric acid;-   AT 4-(4-(2,5-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid;-   AU 4-(4-(2,5-Difluorobenzyloxy)phenyl)-4-oxobutyric acid;-   AV 4-(4-(2,4-Difluorobenzyloxy)phenyl)-4-oxobutyric acid;-   AW 4-(3-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyric acid;-   AX 4-(4-((Cyclopropyl)-methoxy)phenyl)-4-oxobutyric acid;-   AY 4-(4-(2-Trifluoromethylbenzyloxy)phenyl)-4-oxobutyric acid;-   AZ 3-[(4-(2,6-Difluorobenzyloxy)phenyl)-methylthio]propionic acid;-   BA 4-(2-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyric acid;-   BB Ethyl 4-(4-(2,6-difluorobenzyloxy)phenyl)-3-oxobutyrate;-   BC    3-(2-(4-(2,6-Difluorobenzyloxy)phenyl)-2-oxoethyl)thio-1H-1,2,4-triazole;-   BD 5-[(4-(2,6-Difluorobenzyloxy)phenyl)-methyl]-1H-tetrazole;-   BE    (2RS)2-(N-Boc)-3-[2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl]thiopropionic    acid;-   BF Ethyl    2-Hydroxy-4-oxo-4-(4-(2,6-difluorobenzyloxy)phenyl)but-2-enoate;-   BG    (2RS)2-(N-Acetyl)-4-(4-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyric    acid;-   BH 4-(3-((Cyclopropyl)-methoxy)phenyl)-4-oxobutyric acid;-   BI 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid;-   BJ 4-(3-(2-Fluoro-6-methylbenzyloxy)phenyl)-4-oxobutyric acid;-   BK Ethyl 4-(3-(2,6-dimethylbenzyloxyl)phenyl)-4-oxobutyrate;-   BL 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid Sodium    salt;-   BM 4-(4-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid;-   BN 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid Potassium    salt;-   BO 4-(3-(2,6-Dimethoxybenzyloxy)phenyl)-4-oxobutyric acid;-   BP 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxo-2,2-dimethylbutyric    acid;-   BQ 4-(3-(4-Trifluoromethylbenzyloxy)phenyl)-4-oxobutyric acid;-   BR 4-(3-((Cyclobutyl)-methoxy)phenyl)-4-oxobutyric acid;-   BS 4-(3-(2,6-Dimethylbenzyloxy)phenyl)butyric acid;-   BT 4-[[4-(2,6-Dimethylbenzyloxy)-3-methoxy]phenyl]-4-oxobutyric    acid;-   BU    4-{3-[((4-Trifluoromethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyric    acid;-   BV    4-{3-[((2,6-Dimethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyric    acid;-   BW 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutanecarbohydroxamic    acid;-   BX 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyramide;-   BY 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxo-2-butenoic acid; and-   BZ 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-3-butenoic acid.

As used herein the transitional term “comprising” is open-ended. A claimutilizing this term can contain elements in addition to those recited insuch claim.

Detailed Description of Active Compounds

In an embodiment of the agent of Formula I′, the agent is a compound ofthe formula:

wherein n is 1 or 2; m is 0 or 1; q is 0 or 1; t is 0 or 1; R⁵ is alkylhaving from 1 to 3 carbon atoms; A is phenyl, unsubstituted orsubstituted by 1 or 2 groups selected from: halo, alkyl having 1 or 2carbon atoms, perfluoromethyl, alkoxy having 1 or 2 carbon atoms, andperfluoromethoxy; or cycloalkyl having from 3 to 6 ring carbon atomswherein the cycloalkyl is unsubstituted or one or two ring carbons areindependently mono-substituted by methyl or ethyl; or a 5 or 6 memberedheteroaromatic ring having 1 or 2 ring heteroatoms selected from N, Sand O and the heteroaromatic ring is covalently bound to the remainderof the compound of formula I by a ring carbon; and X is —CH₂— and R¹ isethyl; or X is —CH₂CH₂— or —CH₂CH(NHAc)— and R¹ is hydrogen or alkylhaving from 1 to 7 carbon atoms; or when R¹ is hydrogen, apharmaceutically acceptable salt of the compound.

In different embodiments of the agent of Formula I, R¹ is hydrogen orethyl; q is 0; or X is —CH₂CH₂—.

In another embodiment of the agent of Formula I, A is phenyl,unsubstituted or substituted by 1 or 2 groups selected from: halo, alkylhaving 1 or 2 carbon atoms, perfluoromethyl, alkoxy having 1 or 2 carbonatoms, and perfluoromethoxy each halo is independently fluoro or chloro.In a specific embodiment each halo substituent on phenyl ring A isfluoro. In a more specific embodiment phenyl ring A is substituted by 2fluoro groups. In a specific embodiment the alkyl, perluoroalkyl, alkoxyor perfluoroalkoxy has 1 carbon atom.

In another embodiment of the agent of Formula I, A is cycloalkyl havingfrom 3 to 6 ring carbon atoms wherein the cycloalkyl is unsubstituted orone or two ring carbons are independently mono-substituted by methyl orethyl. In a specific embodiment the cycloalkyl is unsubstituted or oneor both ring carbons adjacent to the ring carbon covalently bound to theremainder of the compound of formula I are independentlymono-substituted by methyl or ethyl. In a more specific embodiment A isunsubstituted cyclopropyl.

In another embodiment of the agent of Formula I, q is 1 and R⁵ ismethyl.

In another embodiment the agent is a compound of the formula:

wherein n is 1 or 2; m is 0 or 1; q is 0 or 1; t is 0 or 1; R² and R³selected from hydrogen, halo, alkyl having 1 or 2 carbon atoms,perfluoromethyl, alkoxy having 1 or 2 carbon atoms, andperfluoromethoxy; R⁵ is alkyl having from 1 to 3 carbon atoms; and X is—CH₂— and R¹ is ethyl; or X is —CH₂CH₂— or —CH₂CH(NHAc)— and R¹ ishydrogen or alkyl having from 1 to 7 carbon atoms; or when R¹ ishydrogen, a pharmaceutically acceptable salt of the compound. In a morespecific embodiment R¹ is hydrogen or ethyl. Examples of compounds ofFormula IA include Compound AM and Compound BG.

In a specific embodiment the agent is a compound of the formula:

wherein n is 1 or 2; m is 0 or 1; p is 1 and R¹ is ethyl; or p is 2 andR¹ is hydrogen or alkyl having from 1 to 7 carbon atoms; R² and R³ areeach independently selected from hydrogen, halo, alkyl having 1 or 2carbon atoms, perfluoromethyl, alkoxy having 1 or 2 carbon atoms, andperfluoromethoxy; or when R¹ is hydrogen, a pharmaceutically acceptablesalt of the compound. In a more specific embodiment R¹ is hydrogen orethyl. In a still more specific embodiment one of R² and R³ is hydrogenor halo and the other is halo. Examples of such compounds includeCompound AD, Compound AE and Compound AI. In another still more specificembodiment R² is fluoro and R³ is hydrogen. Examples of such compoundsinclude Compound AA, Compound AJ, Compound AK, and Compound AO. Inanother still more specific embodiment R² is fluoro and R³ is fluoro.Examples of such compounds include Compound AU, Compound AV and CompoundBB.

In a more specific embodiment the agent is a compound of the formula:

wherein n is 1 or 2; m is 0; R¹ is H or alkyl having from 1 to 7 carbonatoms; or when R¹ is hydrogen, a pharmaceutically acceptable salt of thecompound. Examples of such compounds include Compound AH, Compound AQ,Compound AW and Compound BA. In a still more specific embodiment one ofR² and R³ is methyl, methoxy or perfluoromethyl and the other ishydrogen or methyl. In one embodiment R² is methyl, methoxy orperfluoromethyl and R³ is hydrogen. Examples of such compounds includeCompound AB, Compound AL, Compound AN, Compound AP and Compound AY. Inanother embodiment R² is methyl and R³ is methyl. Examples of suchcompounds include Compound AT and Compound BI. In another embodiment R²is hydrogen and R³ is hydrogen. Examples of such compounds includeCompound AG.

In another embodiment the agent is a compound of the formula:

wherein R¹ is hydrogen or alkyl having from 1 to 7 carbon atoms, or whenR¹ is hydrogen, a pharmaceutically acceptable salt of the compound. In aspecific embodiment R¹ is hydrogen or ethyl. Examples of such compoundsinclude Compound AX and Compound BH. In another embodiment the agent isa compound of the formula:

wherein n is 1 or 2; R¹ is hydrogen or alkyl having from 1 to 7 carbonatoms; and Het is a 5 or 6 membered heteroaromatic ring having 1 or 2ring heteroatoms selected from N, S and 0 and the heteroaromatic ring iscovalently bound to the remainder of the compound of formula IC by aring carbon. In a specific embodiment R¹ is hydrogen or ethyl. Examplesof such compounds include Compound AF and Compound AR.

In an embodiment of the agent of Formula II, A is cycloalkyl having from3 to 6 ring carbon atoms wherein the cycloalkyl is unsubstituted or oneor both of the ring carbons adjacent to the remainder of the compound offormula II are mono-substituted by methyl or ethyl. In anotherembodiment of the agent of Formula II, A is phenyl, unsubstituted orsubstituted by 1 or 2 groups selected from: fluoro, alkyl having 1 or 2carbon atoms, perfluoromethyl, alkoxy having 1 or 2 carbon atoms, andperfluoromethoxy.

In another embodiment, the agent is a compound of the formula:

wherein m is 0 or 1; r is 0 or 1; Z is

R¹ is hydrogen or alkyl having from 1 to 7 carbon atoms; R⁴ is hydrogen;—NHCOOC(CH₃)₃; —NHCH₃; or —NHCH₂CH₃; R³ is hydrogen or halo; or when R¹is hydrogen, a pharmaceutically acceptable salt of the compound. In aspecific embodiment R¹ is hydrogen or ethyl. Examples of such compoundsinclude Compound AC, Compound AZ, Compound BC and Compound BE.

In an embodiment of the agent of Formula III, A is phenyl, unsubstitutedor substituted by 1 or 2 groups selected from: halo, alkyl having 1 or 2carbon atoms, perfluoromethyl, alkoxy having 1 or 2 carbon atoms, andperfluoromethoxy. Examples of such compounds include Compound BD.

In an embodiment of the agent of Formula IV, R¹ is hydrogen or ethyl.Examples of such compounds include Compound AS.

In an embodiment of the agent of Formula V′, the agent is a compound ofthe formula:

wherein n is 1 or 2; R¹ is hydrogen or alkyl having from 1 to 7 carbonatoms; A is phenyl, unsubstituted or substituted by 1 or 2 groupsselected from halo, alkyl having 1 or 2 carbon atoms, perfluoromethyl,alkoxy having 1 or 2 carbon atoms, and perfluoromethoxy; or cycloalkylhaving from 3 to 6 ring carbon atoms wherein the cycloalkyl isunsubstituted or one or two ring carbons are independentlymono-substituted by methyl or ethyl; or a 5 or 6 membered heteroaromaticring having 1 or 2 ring heteroatoms selected from N, S and O and theheteroaromatic ring is covalently bound to the remainder of the compoundof formula V by a ring carbon; or a pharmaceutically acceptable salt ofthe compound.

In an embodiment of the agent of Formula V, the agent is a compound ofthe formula:

wherein n is 1 or 2; R¹ is hydrogen or alkyl having from 1 to 7 carbonatoms; R² and R³ are each independently selected from hydrogen, halo,alkyl having 1 or 2 carbon atoms, perfluoromethyl, alkoxy having 1 or 2carbon atoms, and perfluoromethoxy, or a pharmaceutically acceptablesalt of the compound. In a specific embodiment R¹ is hydrogen or ethyl.Examples of such compounds include Compound BF.Use in Methods of Treatment

This invention provides a method for treating a mammalian subject with acondition selected from the group consisting of insulin resistancesyndrome and diabetes (both primary essential diabetes such as Type IDiabetes or Type II Diabetes and secondary nonessential diabetes),comprising administering to the subject an amount of a biologicallyactive agent as described herein effective to treat the condition. Inaccordance with the method of this invention a symptom of diabetes orthe chance of developing a symptom of diabetes, such as atherosclerosis,obesity, hypertension, hyperlipidemia, fatty liver disease, nephropathy,neuropathy, retinopathy, foot ulceration and cataracts, each suchsymptom being associated with diabetes, can be reduced. This inventionalso provides a method for treating hyperlipidemia comprisingadministering to the subject an amount of a biologically active agent asdescribed herein effective to treat the condition. As shown in theExamples, compounds reduce serum triglycerides and free fatty acids inhyperlipidemic animals. This invention also provides a method fortreating cachexia comprising administering to the subject an amount of abiologically active agent as described herein effective to treat thecachexia. This invention also provides a method for treating obesitycomprising administering to the subject an amount of a biologicallyactive agent as described herein effective to treat the condition. Thisinvention also provides a method for treating a condition selected fromatherosclerosis or arteriosclerosis comprising administering to thesubject an amount of a biologically active agent as described hereineffective to treat the condition. The active agents of this inventionare effective to treat hyperlipidemia, fatty liver disease, cachexia,obesity, atherosclerosis or arteriosclerosis whether or not the subjecthas diabetes or insulin resistance syndrome. The agent can beadministered by any conventional route of systemic administration.Preferably the agent is administered orally. Other routes ofadministration that can be used in accordance with this inventioninclude rectally, parenterally, by injection (e.g. intravenous,subcutaneous, intramuscular or intraperitioneal injection), or nasally.

Further embodiments of each of the uses and methods of treatment of thisinvention comprise administering any one of the embodiments of thebiologically active agents described above. In the interest of avoidingunnecessary redundancy, each such agent and group of agents is not beingrepeated, but they are incorporated into this description of uses andmethods of treatment as if they were repeated.

Many of the diseases or disorders that are addressed by the compounds ofthe invention fall into two broad categories: Insulin resistancesyndromes and consequences of chronic hyperglycemia. Dysregulation offuel metabolism, especially insulin resistance, which can occur in theabsence of diabetes (persistent hyperglycemia) per se, is associatedwith a variety of symptoms, including hyperlipidemia, atherosclerosis,obesity, essential hypertension, fatty liver disease (NASH; nonalcoholicsteatohepatitis), and, especially in the context of cancer or systemicinflammatory disease, cachexia. Cachexia can also occur in the contextof Type I Diabetes or late-stage Type II Diabetes. By improving tissuefuel metabolism, active agents of the invention are useful forpreventing or amelioriating diseases and symptoms associated withinsulin resistance, as is demonstrated in animals in the Examples. Whilea cluster of signs and symptoms associated with insulin resistance maycoexist in an individual patient, it many cases only one symptom maydominate, due to individual differences in vulnerability of the manyphysiological systems affected by insulin resistance. Nonetheless, sinceinsulin resistance is a major contributor to many disease conditions,drugs which address this cellular and molecular defect are useful forprevention or amelioration of virtually any symptom in any organ systemthat may be due to, or exacerbated by, insulin resistance.

When insulin resistance and concurrent inadequate insulin production bypancreatic islets are sufficiently severe, chronic hyperglycemia occurs,defining the onset of Type II diabetes mellitus (NIDDM). In addition tothe metabolic disorders related to insulin resistance indicated above,disease symptoms secondary to hyperglycemia also occur in patients withNIDDM. These include nephropathy, peripheral neuropathy, retinopathy,microvascular disease, ulceration of the extremities, and consequencesof nonenzymatic glycosylation of proteins, e.g. damage to collagen andother connective tissues. Attenuation of hyperglycemia reduces the rateof onset and severity of these consequences of diabetes. Because, as isdemonstrated in the Examples, active agents and compositions of theinvention help to reduce hyperglycemia in diabetes, they are useful forprevention and amelioration of complications of chronic hyperglycemia.

Both human and non-human mammalian subjects can be treated in accordancewith the treatment method of this invention. The optimal dose of aparticular active agent of the invention for a particular subject can bedetermined in the clinical setting by a skilled clinician. In the caseof oral administration to a human for treatment of disorders related toinsulin resistance, diabetes, hyperlipidemia, fatty liver disease,cachexia or obesity the agent is generally administered in a daily doseof from 1 mg to 400 mg, administered once or twice per day. For oraladministration to a human the anticipated preferred daily dose ofCompound AH is from 100 mg to 400 mg; of Compound AW is from 30 to 300mg; and of Compound BI is from 10 to 200 mg. In the case of oraladministration to a mouse the agent is generally administered in a dailydose from 1 to 300 mg of the agent per kilogram of body weight. Activeagents of the invention are used as monotherapy in diabetes or insulinresistance syndrome, or in combination with one or more other drugs withutility in these types of diseases, e.g. insulin releasing agents,prandial insulin releasers, biguanides, or insulin itself. Suchadditional drugs are administered in accord with standard clinicalpractice. In some cases, agents of the invention will improve theefficacy of other classes of drugs, permitting lower (and therefore lesstoxic) doses of such agents to be administered to patients withsatisfactory therapeutic results. Established safe and effective doseranges in humans for representative compounds are: metformin 500 to 2550mg/day; glyburide 1.25 to 20 mg/day; GLUCOVANCE (combined formulation ofmetformin and glyburide) 1.25 to 20 mg/day glyburide and 250 to 2000mg/day metformin; atorvastatin 10 to 80 mg/day; lovastatin 10 to 80mg/day; pravastatin 10 to 40 mg/day; and simvastatin 5-80 mg/day;clofibrate 2000 mg/day; gemfibrozil 1200 to 2400 mg/day, rosiglitazone 4to 8 mg/day; pioglitazone 15 to 45 mg/day; acarbose 75-300 mg/day;repaglinide 0.5 to 16 mg/day.

Type I Diabetes Mellitus: A patient with Type I diabetes manages theirdisease primarily by self-administration of one to several doses ofinsulin per day, with frequent monitoring blood glucose to permitappropriate adjustment of the dose and timing of insulin administration.Chronic hyperglycemia leads to complications such as nephropathy,neuropathy, retinopathy, foot ulceration, and early mortality;hypoglycemia due to excessive insulin dosing can cause cognitivedysfunction or unconsciousness. A patient with Type I diabetes istreated with 1 to 400 mg/day of an active agent of this invention, e.g.50 to 400 mg/day of Compound AH, in tablet or capsule form either as asingle or a divided dose. The anticipated effect will be a reduction inthe dose or frequency of administration of insulin required to maintainblood glucose in a satisfactory range, and a reduced incidence andseverity of hypoglycemic episodes. Clinical outcome is monitored bymeasurement of blood glucose and glycosylated hemoglobin (an index ofadequacy of glycemic control integrated over a period of severalmonths), as well as by reduced incidence and severity of typicalcomplications of diabetes. A biologically active agent of this inventioncan be administered in conjunction with islet transplantation to helpmaintain the anti-diabetic efficacy of the islet transplant.

Type II Diabetes Mellitus: A typical patient with Type II diabetes(NIDDM) manages their disease by programs of diet and exercise as wellas by taking medications such as metformin, glyburide, repaglinide,rosiglitazone, or acarbose, all of which provide some improvement inglycemic control in some patients, but none of which are free of sideeffects or eventual treatment failure due to disease progression. Isletfailure occurs over time in patients with NIDDM, necessitating insulininjections in a large fraction of patients. It is anticipated that dailytreatment with an active agent of the invention (with or withoutadditional classes of antidiabetic medication) will improve glycemiccontrol, reduce the rate of islet failure, and reduce the incidence andseverity of typical symptoms of diabetes. In addition, active agents ofthe invention will reduce elevated serum triglycerides and fatty acids,thereby reducing the risk of cardiovascular disease, a major cause ofdeath of diabetic patients. Suitable daily dose ranges for selectedcompounds of the invention for treatment of NIDDM (either as monotherapyor in combination with other antidiabetic drugs) are from 50 mg to 400mg of Compound AH, from 15 mg to 300 mg of Compound AW, or from 5 mg to200 mg of Compound BI. As is the case for all other therapeutic agentsfor diabetes, dose optimization is done in individual patients accordingto need, clinical effect, and susceptibility to side effects.

Hyperlipidemia: Elevated triglyceride and free fatty acid levels inblood affect a substantial fraction of the population and are animportant risk factor for atherosclerosis and myocardial infarction.Active agents of the invention are useful for reducing circulatingtriglycerides and free fatty acids in hyperlipidemic patients. Suitabledaily dose ranges for selected compounds of the invention for treatmentof hypertriglyceridemia are from 50 mg to 400 mg of Compound AH, from 15mg to 300 mg of Compound AW, or from 5 mg to 200 mg of Compound BI.Hyperlipidemic patients often also have elevated blood cholesterollevels, which also increase the risk of cardiovascular disease.Cholesterol-lowering drugs such as HMG-CoA reductase inhibitors(“statins”) can be administered to hyperlipidemic patients in additionto agents of the invention, optionally incorporated into the samepharmaceutical composition.

Fatty Liver Disease: A substantial fraction of the population isaffected by fatty liver disease, also known as nonalcoholicsteatohepatitis (NASH); NASH is often associated with obesity anddiabetes. Hepatic steatosis, the presence of droplets of triglycerideswith hepatocytes, predisposes the liver to chronic inflammation(detected in biopsy samples as infiltration of inflammatory leukocytes),which can lead to fibrosis and cirrhosis. Fatty liver disease isgenerally detected by observation of elevated serum levels ofliver-specific enzymes such as the transaminases ALT and AST, whichserve as indices of hepatocyte injury, as well as by presentation ofsymptoms which include fatigue and pain in the region of the liver,though definitive diagnosis often requires a biopsy. As is shown in theExamples, compounds of the invention, e.g. Compound AW, reduce serumliver transaminases and liver fat content in an established animal modelof NASH (ob/ob obese mice), and are therefore useful for treatment offatty liver disease. An appropriate dose range of Compound AW fortreatment of fatty liver disease is 15 to 300 mg/day. The anticipatedbenefit is a reduction in liver inflammation and fat content, resultingin attenuation, halting, or reversal of the progression of NASH towardfibrosis and cirrhosis.

Pharmaceutical Compositions

This invention provides a pharmaceutical composition comprising abiologically active agent as described herein and a pharmaceuticallyacceptable carrier. Further embodiments of the pharmaceuticalcomposition of this invention comprise any one of the embodiments of thebiologically active agents described above. In the interest of avoidingunnecessary redundancy, each such agent and group of agents is not beingrepeated, but they are incorporated into this description ofpharmaceutical compositions as if they were repeated.

Preferably the composition is adapted for oral administration, e.g. inthe form of a tablet, coated tablet, dragee, hard or soft gelatincapsule, solution, emulsion or suspension. In general the oralcomposition will comprise from 1 mg to 400 mg of such agent. It isconvenient for the subject to swallow one or two tablets, coatedtablets, dragees, or gelatin capsules per day. Accordingly, preferredoral compositions for treatment of humans comprise from 50 mg to 400 mgof Compound AH, from 15 mg to 300 mg of Compound AW, or from 5 mg to 200mg of Compound BI. However the composition can also be adapted foradministration by any other conventional means of systemicadministration including rectally, e.g. in the form of suppositories,parenterally, e.g. in the form of injection solutions, or nasally.

The biologically active compounds can be processed with pharmaceuticallyinert, inorganic or organic carriers for the production ofpharmaceutical compositions. Lactose, corn starch or derivativesthereof, talc, stearic acid or its salts and the like can be used, forexample, as such carriers for tablets, coated tablets, dragees and hardgelatin capsules. Suitable carriers for soft gelatin capsules are, forexample, vegetable oils, waxes, fats, semi-solid and liquid polyols andthe like. Depending on the nature of the active ingredient no carriersare, however, usually required in the case of soft gelatin capsules,other than the soft gelatin itself. Suitable carriers for the productionof solutions and syrups are, for example, water, polyols, glycerol,vegetable oils and the like. Suitable carriers for suppositories are,for example, natural or hardened oils, waxes, fats, semil-liquid orliquid polyols and the like.

The pharmaceutical compositions can, moreover, contain preservatives,solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners,colorants, flavorants, salts for varying the osmotic pressure, buffers,coating agents or antioxidants. They can also contain still othertherapeutically valuable substances, particularly antidiabetic orhypolipidemic agents that act through mechanisms other than thoseunderlying the effects of the compounds of the invention. Agents whichcan advantageously be combined with compounds of the invention in asingle formulation include but are not limited to biguanides such asmetformin, insulin releasing agents such as the sulfonylurea insulinreleaser glyburide and other sulfonylurea insulin releasers,cholesterol-lowering drugs such as the “statin” HMG-CoA reductaseinhibitors such as atrovastatin, lovastatin, pravastatin andsimvastatin, PPAR-alpha agonists such as clofibrate and gemfibrozil,PPAR-gamma agonists such as thiazolidinediones (e.g. rosiglitazone andpioglitazone, alpha-glucosidase inhibitors such as acarbose (whichinhibit starch digestion), and prandial insulin releasers such asrepaglinide. The amounts of complementary agents combined with compoundsof the invention in single formulations are in accord with the dosesused in standard clinical practice. Established safe and effective doseranges for certain representative compounds are set forth above.

Reaction Schemes

The biologically active compounds of the present invention can be madein accordance with the following reaction schemes.

The compound of formula I′ where X is —CH₂CR¹²R¹³—, q and m are 0, t is0 or 1, and n is 1 or 2, R⁹ is hydrogen, halo, or alkoxy having 1 to 3carbon atoms, Q is OR¹ where R¹ is hydrogen or alkyl having from 1 to 7carbons, i.e. compounds of formula:

wherein A is as described above, and R¹ is hydrogen or alkyl having from1 to 7 carbon atoms, R¹² and R¹³ is independently hydrogen or methyl canbe prepared from the compound of formula VI via the reaction scheme inScheme 1.

In the reaction scheme of Scheme 1, A, t, n and R⁹ are as above. R⁶ isan alkyl group containing from 1 to 7 carbon atoms, R¹² and R¹³ isindependently hydrogen or methyl and Y is a leaving group.

The compound of formula VI is converted to the compound of formula VIIIvia reaction of step (a) using Mitsunobu condensation of VI with VIIusing triphenylphosphine and diethyl azodicarboxylate. Any of theconditions conventionally used in Mitsunobu reactions can be utilized tocarry out the reaction of step (a).

The compound of formula VIII can also be prepared by etherifying oralkylating the compound of formula VI with a compound of formula IX asin reaction of step (b). In the compound of formula IX, Y can be anyconventional leaving group such as mesyloxy, tosyloxy or a halide. Anyconventional method of etherifying of a hydroxyl group through reactionwith a halide or leaving group can be utilized to carry out the reactionof step (b). The reaction of step (b) is preferred over step (a) ifcompound of formula IX is readily available.

The compound of formula VIII is converted to the compound of formula XIvia reaction of step (c) by alkylating the compound of formula VIII withthe compound of formula X. This reaction is carried out utilizing aconventional base which converts acetophenone to 3-keto ester (i.e.gamma-keto ester). Any conventional base for this purpose can beutilized in the reaction of step (c). In carrying out this reaction itis generally preferred to utilize alkali metal salts ofhexamethyldisilazane such as lithium bis(trimethylsilyl)amide as base.Generally this reaction is carried out in an inert solvent such astetrahydrofuran: 1,3-Dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone(5:1). Any of the conditions conventional in such alkylation reactionscan be utilized to carry out the reaction of step (c).

The compound of formula XI is the compound of formula I′ where R¹ is analkyl group containing from 1 to 7 carbon atoms. The compound of formulaXI can be converted to the free acid i.e. the compound of formula I′where R¹ is H by ester hydrolysis. Any conventional method of esterhydrolysis will produce the compound of formula I′ where R¹ is H.

The compound of general formula VII can be prepared by reducing thecorresponding acid of formula A-(CH₂)_(t+n)—CO₂H. The reaction iscarried out first by esterification of compound of formulaA-(CH₂)_(t+n)CO₂H with methyl iodide, followed by reduction utilizing aconventional base for example, lithium aluminium hydride or the like inan inert organic solvent for example, tetrahydrofuran or the like. Anyof the conditions conventional in such reduction reactions can beutilized to carry out this reaction.

The compound of formula VII where A is 2,6 Dimethyl phenyl can beprepared from the compound of formula XCI, via the reaction scheme inScheme 2.

In Scheme 2, the compound of formula XCI can be converted to compound offormula VII by esterification with methyl iodide, followed by reductionwith lithium aluminum hydride via reaction of step (r″). The reaction ofstep (r″) can be carried out utilizing a conventional reducing agent. Incarrying out this reaction it is generally preferred to utilize lithiumaluminum hydride as the reducing agent. Any of the conditionsconventional in reduction reactions can be utilized to carry out thisreaction.

The compound of formula I where X is —CH₂—, q is 0, m is 1, t is 0 or 1and n is 1 or 2, i.e. compounds of the formula:

wherein A is as described above, R¹ is ethyl, and R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms can be prepared from the compoundof formula XII, wherein m is as above via the reaction scheme in Scheme3.

In Scheme 3, A is as above, Y is a leaving group such as halide,mesyloxy or tosyloxy. Y¹ is chloro.

In Scheme 3, the compound of formula XII is converted into the ethylester of formula XIII using ethanol via reaction of step (d). Anyconventional method of converting acid to ethyl ester can be utilized tocarry out this reaction.

The compound of formula XIII can be converted to compound of formula XIVin the same manner as described in the connection with reaction of step(a) or (b) hereinbefore.

In the step of (f), the compound of formula XIV is hydrolyzed to producethe compound of formula XV. Any conventional method of basic hydrolysisto hydrolyze ester can be utilized to carry out this reaction.

The compound of formula XV is converted to the acid chloride of formulaXVI via reaction of step (g) by reaction with thionyl chloride. Any ofthe conventional methods of converting acid to acid halide can beutilized to carry out this reaction of step (g).

The compound of formula XVII is reacted with acid chloride of formulaXVI to produce the compound of formula XVIII via reaction of step (h).Any conventional base can be utilized to carry out this reaction withthe preferred base being pyridine. The resulting acylated Meldrum acidswere not isolated, and instead after workup they were refluxed inabsolute ethanol to give the 2-ketoesters. Any conventional conditionsto carry out the reaction of step (h) can be utilized.

The compound of formula XVIII is the compound of formula I where R¹ isethyl.

The compound of formula I′ where q is 1, R⁵ is an alkyl group having 1to 3 carbon atoms where X is —CH₂CR¹²R¹³—, m is 0, t is 0 or 1 and n is1 or 2, i.e. compounds of the formula:

wherein A is as above, R¹ is hydrogen or alkyl having from 1 to 7 carbonatoms, and R¹² and R¹³ is independently hydrogen or methyl, R⁹ ishydrogen, halo, or alkoxy having 1 to 3 carbon atoms, Q is OR¹ where R¹is hydrogen or alkyl having from 1 to 7 carbons, can be prepared fromthe compound of formula XIX, wherein t and A are as above via thereaction scheme in Scheme 4.

In Scheme 4, t, n, A, R¹, R⁹, R¹², R¹³ and R⁵ are as above. R⁶ is analkyl group having 1 to 7 carbon atoms. Y¹ is chloro.

In Scheme 4, the compound of formula XIX is mesylated to furnish thecompound of formula XX via reaction of step (i). Any conventionalconditions to carry out mesylation can be utilized. The compound offormula XX is then heated with the compound of formula XXI to producethe compound of formula XXII. Any of the conditions conventional toproduce amino alcohol can be utilized in reaction of step (j).

In the compound of formula XXII, alcohol is then displaced by chloro bytreating the compound of formula XXII with thionyl chloride to producecompound of formula XXIII via reaction of step (k). Any conventionalmethod to displace alcohol with halo can be utilized to carry out thisreaction.

The compound of formula XXIII is reacted with a compound of formula VIin the presence of base using dimethylformamide as solvent via reactionof step (l) to produce the corresponding compound of formula XXIV. Theposition of the substituents in the compound of formula VI willdetermine the position of the substituents in the compound of formulaXXIV. Any conventional method of etherification of a hydroxyl group inthe presence of base (preferred base being potassium carbonate) with ahalide can be utilized to carry out the reaction of step (l). Thecompound of formula XXIV is converted to the compound of formula XXV viareaction of step (m) by alkylating the compound of formula XXIV with thecompound of formula X in the presence of alkali metal silyl amide asbase (eg. lithium hexamethyldisilane or sodium hexamethyldisilane). Thisreaction is carried out in the same manner as described in connectionwith reaction of step (c) of Scheme 1.

The compound of formula XXV is the compound of formula I′ where R¹ is analkyl group having 1 to 7 carbon atoms. The compound of formula XXV canbe converted to the free acid i.e. the compound of formula I′ where R¹is H by ester hydrolysis. Any conventional method of ester hydrolysiswill produce the compound of formula I′ where R¹ is H.

The compound of formula I′ where X is —CH₂CH(NHAc),— m is 0, q is 0, tis 0 or 1 and n is 1 or 2, i.e. compounds of the formula:

wherein A is as above, R¹ is hydrogen on alkyl having from 1 to 7 carbonatoms, and R⁹ is hydrogen, halo, or alkoxy having 1 to 3 carbon atoms, Qis OR¹ where R¹ is hydrogen or alkyl having from 1 to 7 carbons can beprepared from the compound of formula VIII, via the reaction scheme inScheme 5.

In Scheme 5, t, n, A, R⁹ and R¹ are as above. R⁷ is an alkyl grouphaving 1 to 7 carbon atoms.

The compound of formula VIII is prepared in the same manner as describedhereinbefore in connection with reaction of step (a) or (b) in Scheme 1.

The compound of formula VIII is converted to compound of formula XXVI byselective bromination of the methyl ketone moiety via reaction of step(n) by treating the compound of formula VIII with CuBr₂. Any selectivebromination conditions to convert methyl ketone to 1-bromoketone can beutilized to carry out the reaction of step (n).

The compound of formula XXVI can be converted to compound of formulaXXVIII via reaction of step (o) by treating the compound of formula XXVIwith the sodium salt of compound of formula XXVII in ethanol. Anyconventional conditions for this alkylation reaction can be utilized tocarry out this reaction.

The compound of formula XXVIII is converted to compound of formula XXIXvia reaction of step (p) by de-esterification employing 4 equivalents ofsodium hydroxide.

Initial mono de-esterification followed by slow hydrolysis of theremaining ethyl ester was observed. Removal of solvent and incubation ofthe residue in acetic acid produced the compound of formula XXIX.

The compound of formula XXIX is the compound of formula I′ where R¹ isH.

The compound of formula XXIX can be converted to compound of formulaXXXI where R⁷ is an alkyl chain having 1 to 7 carbon atoms byesterification of carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (q).

The compound of formula XXXI is the compound of formula I′ where R¹ isan alkyl chain having 1 to 7 carbon atoms.

The compound of formula I′ where X is —CH₂—, q and m are 0, t is 0 or 1and n is 1 or 2, i.e. compounds of formula:

wherein t, n, and A are as described above, R⁹ is hydrogen, halo, oralkoxy having 1 to 3 carbon atoms and R¹ is ethyl can be prepared fromthe compound of formula LX, via the reaction scheme in Scheme 6.

In the reaction scheme of Scheme 6, A, t, R⁹ and n are as above, Y is aleaving group and Y¹ is chloro.

In Scheme 6, the compound of formula LX is converted to compound offormula LXI in the same manner as described hereinbefore in connectionwith the reaction of steps (a) or (b) in Scheme 1.

In the step of (q′), the compound of formula LXI is hydrolyzed toproduce the compound of formula LXII in the same manner as described inconnection with the reaction of step (f) in Scheme 3.

The compound of formula LXII is converted to compound of formula LXIIIvia reaction of step (r′) in the same manner as described in connectionwith reaction of step (g) in Scheme 3.

The compound of formula LXIV is first treated with 2 equivalents ofn-butyllithium at low temperature and then compound of formula LXIII isadded to produce compound of formula LXV (Weirenga, W.; Skulnick, H. I.J.O.C. 1979, 44, 310-311). The compound of formula LXV is the compoundof formula 1 where R¹ is ethyl.

The compound of formula I where q is 1, R⁵ is an alkyl group having 1 to3 carbon atoms, where X is —CH₂—, m is 0, t is 0 or 1 and n is 1 or 2,i.e. compounds of formula:

wherein A is as described above, R⁹ is hydrogen, halo, or alkoxy having1 to 3 carbon atoms and R¹ is ethyl can be prepared from the compound offormula LX via the reaction scheme in Scheme 7.

In the reaction scheme of Scheme 7, A, t, R⁹ and n are as above, Y¹ ischloro. R⁵ is an alkyl group having from 1 to 3 carbon atoms.

In Scheme 7, the compound of formula LX is reacted with compound offormula XXIII (prepared in the same manner as described in Scheme 4) toproduce the compound of formula LXVI via reaction of step (t′). Thisreaction is carried out in the same manner as described hereinbefore inthe connection with reaction of step (l) in Scheme 4.

In the step of (u′), the compound of formula LXVI is hydrolyzed toproduce the compound of formula LXVII in the same manner as described inthe reaction of step (f) in Scheme 3.

The compound of formula LXVII is converted to compound of formula LXVIIIvia reaction of step (v′) in the same manner as described in connectionwith the reaction of step (g) in Scheme 3.

The compound of formula LXIV is first treated with 2 equiv ofn-butyllithium at low temperature and then compound of formula LXIII isadded to produce compound of formula LXV (Weirenga, W.; Skulnick, H. I.J.O.C. 1979, 44, 310-311).

The compound of formula LXIX is the compound of formula I′ where R¹ isan alkyl group having 2 carbon atoms.

The compound of formula I′ where q is 1, R⁵ is an alkyl group having 1to 3 carbon atoms, R¹ is hydrogen or alkyl having from 1 to 7 carbons,R⁹ is hydrogen, halo, or alkoxy having 1 to 3 carbon atoms, Q is OR¹where R¹ is hydrogen or alkyl having from 1 to 7 carbons, X is—CH₂CH(NHAc)—, m is 0, t is 0 or 1 and n is 1 or 2, i.e. compounds ofthe formula:

wherein t, n, A and R¹ are as above, can be prepared from the compoundof formula VI, via the reaction scheme in Scheme 8.

In Scheme 8, t, n, A, R⁹ and R¹ are as above. R⁷ is an alkyl grouphaving 1 to 7 carbon atoms. R⁵ is an alkyl group having 1 to 3 carbonatoms. Y¹ is chloro.

The compound of formula XXIV is prepared in the same manner as describedhereinbefore in connection with reaction of step (l) in Scheme 4.

The compound of formula XXIV is converted to compound of formula LXX byselective bromination of the methyl ketone moiety via reaction of step(x′) by treating the compound of formula XXIV with CuBr₂. Any selectivebromination conditions to convert methyl ketone to 1-bromoketone can beutilized to carry out the reaction of step (x′).

The compound of formula LXX can be converted to the compound of formulaLXXI via reaction of step (y′) by treating the compound of formula LXXwith the sodium salt of compound of formula XXVII in ethanol. Anyconventional conditions can be utilized to carry out alkylationreaction.

The compound of formula LXXI is converted to the compound of formulaLXXII via reaction of step (z′) by de-esterification employing 4 equiv.of sodium hydroxide. This indicated an initial mono de-esterificationfollowed by slow hydrolysis of the remaining ethyl ester. Removal ofsolvent and incubation of the residue in acetic acid produced thecompound of formula LXXII.

The compound of formula LXXII is the compound of formula I′ where R¹ isH.

The compound of formula LXXII can be converted to compound of formulaLXXIII where R⁷ is an alkyl group having 1 to 7 carbon atoms byesterification of carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (a″).

The compound of formula LXXIII is the compound of formula I′ where R¹ isan alkyl group having 1 to 7 carbon atoms.

The compound of formula I′ where X is —CH₂CH(NHAc)—, R⁹ is hydrogen,halo, or alkoxy having 1 to 3 carbon atoms, Q is OR¹ where R¹ ishydrogen or alkyl having from 1 to 7 carbons, m is 1, q is 0, t is 0 or1 and n is 1 or 2, i.e. compounds of the formula:

wherein A is as above, and R¹ is hydrogen or alkyl having from 1 to 7carbon atoms can be prepared from the compound of formula LXXIV via thereaction scheme in Scheme 9.

In Scheme 9, t, n, A, R⁹ and R¹ are as above. R⁷ is an alkyl grouphaving 1 to 7 carbon atoms. R⁵ is an alkyl group having 1 to 3 carbonatoms.

The compound of formula LXXIV can be prepared according to methoddescribed in Murphy et al J.C.S. Perkin 1, 1980, 1555-1566.

The compound of formula LXXIV can be alkylated to produce compound offormula LXXV via reaction of step (b″) employing either compound offormula VII using same method as described in the connection of reactionstep of (a) in Scheme 1 or compound of formula IX using potassiumcarbonate as the base for alkylation. The reaction is carried out in thesame manner as described hereinbefore in connection with the reaction ofstep (l) in Scheme 4.

The compound of formula LXXV is then selectively brominated at 0° C.using 30 wt % HBr in acetic acid dropwise to produce compound of formulaLXXVI via reaction of step (c″). Any conventional method to convertselectively substituted acetone to 1-Bromoacetone can be utilized tocarry out this reaction of step (c″).

The compound of formula LXXVI is converted to compound of formula LXXVIIvia reaction of step (d″) in the same manner as described hereinbeforein connection with the reaction of step (o) in Scheme 5.

The compound of formula LXXVII is converted to compound of formulaLXXVIII via reaction of step (e″) by de-esterification employing 4equiv. of sodium hydroxide. Initial mono de-esterification followed byslow hydrolysis of the remaining ethyl ester was observed. Removal ofsolvent and incubation of the residue in acetic acid produced thecompound of formula LXXVIII.

The compound of formula LXXVIII is the compound of formula I′ where R¹is H.

The compound of formula LXXVIII can be converted to compound of formulaLXXIX where R⁷ is an alkyl group having 1 to 7 carbon atoms byesterification of carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (f″).

The compound of formula LXXIX is the compound of formula I′ where R¹ isan alkyl group having 1 to 7 carbon atoms.

The compound of formula I′ where q is 1, R⁵ is an alkyl group having 1to 3 carbon atoms, X is —CH₂CH(NHAc)—, m is 1, t is 0 or 1 and n is 1 or2, R⁹ is hydrogen, halo, or alkoxy having 1 to 3 carbon atoms, Q is OR¹where R¹ is hydrogen or alkyl having from 1 to 7 carbons, i.e. compoundsof the formula:

wherein A is as above, and R¹ is hydrogen or alkyl having from 1 to 7carbon atoms can be prepared from the compound of the formula LXXIV viathe reaction scheme in Scheme 10.

In Scheme 10, t, n, A, R⁹ and R¹ are as above. R⁷ is an alkyl grouphaving 1 to 7 carbon atoms. R⁵ is an alkyl group having 1 to 3 carbonatoms. Y¹ is chloro.

The compound of formula LXXIV can be prepared according to methoddescribed in Murphy et. al. J.C.S. Perkin 1, 1980, 1555-1566.

In Scheme 10, the compound of formula LXXIV is reacted with compound offormula XXIII (prepared in the same manner as described in Scheme 4) toproduce the compound of formula LXXX via reaction of step (g″). Thisreaction is carried out in the same manner as described hereinbefore inconnection with the reaction of step (l) in Scheme 4.

The compound of formula LXXX is then selectively brominated at 0° C.using 30 wt % HBr in acetic acid dropwise to produce compound of formulaLXXXI via reaction of step (h″). Any conventional method to convertsubstituted acetone to 1-Bromoacetone can be utilized to carry out thereaction of step (h″).

The compound of formula LXXXI is converted to the compound of formulaLXXXII via reaction of step (i″) in the same manner as describedhereinbefore in connection with the reaction of step (o) in Scheme 5.

The compound of formula LXXXII is converted to compound of formulaLXXXIII via reaction of step (j″) in the same manner as described inreaction of step (p) in Scheme 5.

The compound of formula LXXXIII is the compound of formula I′ where R¹is H.

The compound of formula LXXXIII can be converted to compound of formulaLXXXIV where R⁷ is an alkyl chain having 1 to 7 carbon atoms byesterification of carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (k″).

The compound of formula LXXXIV is the compound of formula I′ where R¹ isan alkyl group having 1 to 7 carbon atoms.

The compound of formula I′ where X is —CH₂CR¹²R¹³—, R⁹ is hydrogen,halo, or alkoxy having 1 to 3 carbon atoms, Q is OR¹ where R¹ ishydrogen or alkyl having from 1 to 7 carbons, q is 0, m is 1, t is 0 or1 and n is 1 or 2, i.e. compounds of formula:

wherein A is as described above, R¹ is hydrogen or alkyl having from 1to 7 carbon atoms, and R¹² and R¹³ is independently hydrogen or methylcan be prepared from the compound of the formula LXXIV, via the reactionscheme in Scheme 11.

In the reaction scheme of Scheme 11, A, t, R⁹, R¹², R¹³ and n are asabove. R⁶ is an alkyl group containing from 1 to 7 carbon atoms, and Yis a leaving group.

The compound formula LXXV is produced from compound of formula LXXIV inthe same manner as described hereinbefore in connection with thereaction of step (b″) in Scheme 9.

The compound of formula LXXV is converted to compound of formula LXXXVvia reaction of step (l″) by selectively alkylating the compound offormula LXXV with the compound of formula X. This reaction is carriedout utilizing a conventional base which converts substituted ketone togamma-keto ester. In carrying out this reaction it is generallypreferred to utilize lithium diisopropylamide as base. Alkylation willoccur at the less hindered methyl group. Generally this reaction iscarried out in an inert solvent such as tetrahydrofuran or1,2-dimethoxyethane at −78° C.

The compound of formula LXXXV is the compound of formula I′ where R¹ isan alkyl group containing from 1 to 7 carbon atoms. The compound offormula LXXXV can be converted to the free acid i.e. the compound offormula I′ where R¹ is H by ester hydrolysis. Any conventional method ofester hydrolysis will produce the compound of formula I′ where R¹ is H.

The compound of formula I′ where q is 1, R⁵ is an alkyl group having 1to 3 carbon atoms where X is —CH₂—, m is 1, t is 0 or 1 and n is 1 or 2,i.e. compounds of the formula:

wherein A is as above, R⁹ is hydrogen, halo, or alkoxy having 1 to 3carbon atoms and R¹ is ethyl can be prepared from the compound formulaXIII wherein, m is as above via the reaction scheme in Scheme 12.

In Scheme 12, A is as above. Y¹ is chloro.

The compound of formula XIII (prepared in the same manner as describedhereinbefore in connection with the reaction of step (d) in Scheme 3)can be converted to compound of formula LXXXVI via reaction of step (m″)in the same manner as described in the connection with reaction of step(l) in Scheme 4 hereinbefore.

In the step of (n″), the compound of formula LXXXVI is hydrolyzed toproduce the compound of formula LXXXVII. Any conventional method ofbasic hydrolysis to hydrolyze ester can be utilized to carry out thisreaction.

The compound of formula LXXXVII is converted to acid chloride of formulaLXXXVIII via reaction of step (o″) by reaction with thionyl chloride.Any of the conventional method of converting acid to acid halide can beutilized to carry out the reaction.

The compound of formula XVII is reacted with the compound of formulaLXXXVIII to produce the compound of formula LXXXIX via reaction of step(p″). Any conventional base can be used to carry out this reaction withthe preferred base being pyridine. Any conventional conditions to carryout the reaction of step (p″) can be utilized.

The compound of formula LXXXIX is the compound of formula I where R¹ isethyl.

The compound of formula I′ where q is 1, R⁵ is an alkyl group having 1to 3 carbon atoms, where X is —CH₂CR¹²R¹³—, R⁹ is hydrogen, halo, oralkoxy having 1 to 3 carbon atoms, Q is OR¹ where R¹ is hydrogen oralkyl having from 1 to 7 carbons, m is 1, t is 0 or 1 and n is 1 or 2,i.e. compounds of formula:

wherein A is as described above, R¹ is hydrogen or alkyl having from 1to 7 carbon atoms, and R¹² and R¹³ is independently hydrogen or methylcan be prepared from the compound of the formula LXXIV, via the reactionscheme in Scheme 13.

In the reaction scheme of Scheme 13, R⁹, R¹², R¹³, R⁵, A, t, and n areas above. R⁶ is an alkyl group containing from 1 to 7 carbon atoms.

The compound formula LXXX is produced from compound of formula LXXIV inthe same manner as described hereinbefore in connection with thereaction of step (g″) in Scheme 10.

The compound of formula LXXX is converted to compound of formula XC viareaction of step (q″) by alkylating the compound of formula LXXX withthe compound of formula X. This reaction is carried out utilizing aconventional base which converts ketone to 3-keto ester. In carrying outthis reaction it is generally preferred to utilize lithiumdiisopropylamide as base. Alkylation will occur at the less hinderedmethyl group. Generally this reaction is carried out in an inert solventsuch as tetrahydrofuran or 1,2-dimethoxyethane at −78° C.

The compound of formula XC is the compound of formula I′ where R¹ is analkyl group containing from 1 to 7 carbon atoms. The compound of formulaXC can be converted to the free acid i.e. the compound of formula I′where R¹ is H by ester hydrolysis. Any conventional method of esterhydrolysis will produce the compound of formula I′ where R¹ is H.

The compound of formula II where Z is

m is 0, r is 1, q is 0, t is 0 or 1 and n is 1 or 2, R⁴ is—NHCO₂C(CH₃)₃, or —NHCH₂CH₃, i.e. the compounds of formula:

wherein A and R¹ are as above, can be prepared from the compound offormula XXVI via reaction scheme in Scheme 14.

In Scheme 14, t, n, A and R¹ are as above. R⁷ is an alkyl group having 1to 7 carbon atoms. R⁸ is an alkyl group containing from 1 to 2 carbonatoms. Y¹ is halo preferably bromo. In Scheme 14, the compound offormula XXVI (prepared in the same manner as described hereinbefore inconnection with reaction of step (n) in Scheme 5) is reacted with thecompound of formula XXXII in the presence of a base to produce thecompound of formula XXXIII via reaction of step (r). In carrying outthis reaction it is generally preferred to utilize triethylamine asbase. Any conventional method of reacting Boc-cys-OEt with halide can beutilized to carry out this reaction.

The compound of formula XXXIII is compound of formula II where R⁴ is—NHCO₂C(CH₃)₃ and R¹ is ethyl.

The compound of formula XXXIII can be converted to the free acid i.e.the compound of formula II where R¹ is H by ester hydrolysis. Anyconventional method of ester hydrolysis will produce the compound offormula II where R¹ is H and R⁴ is —NHCO₂C(CH₃)₃. The compound offormula XXXIII is converted to compound of formula XXXV first viareaction of step (s) by deprotecting t-butoxy group usingtrifluoroacetic acid and replacing by lower alkyl having 1 to 2 carbonatoms via reaction of step (t). Any conventional method to condenseamine with alkyl halide can be used to carry out this reaction.

The compound of formula XXXV is compound of formula II where R⁴ is anamine having 1 to 2 carbon atoms and R¹ is an alkyl group having 2carbon atoms. The compound of formula XXXV can be converted to the freeacid i.e. the compound of formula XXXVI where R¹ is H by basichydrolysis via reaction of step (u). The compound of formula XXXVI iscompound of formula II where R⁴ is —NHCH₃ or —NHCH₂CH₃ and R¹ is H.

The compound of formula XXXVI can be converted to compound of formulaXXXVII where R⁷ is an alkyl group having 1 to 7 carbon atoms byesterification of carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (v).

The compound of formula XXXVII is compound of formula II where R¹ is analkyl having 1 to 7 carbon atoms and R⁴ is —NHCH₃ or —NHCH₂CH₃.

The compound of formula II where Z is

m and q are 0, r is 1, t is 0 or 1, n is 1 or 2, i.e. compounds of theformula:

A is as above, can be prepared from the compound of formula VIII,wherein t, n and A are as above, via the reaction scheme in Scheme 15.

In Scheme 15, the compound of formula VIII (prepared in the same manneras described hereinbefore in connection with reaction of step (a) or (b)in Scheme 1) is converted to compound of formula XXVI in the same manneras described in reaction of step (n) in Scheme 5.

The compound of formula XXVI is reacted with compound of formula XXXVIIIin the presence of base preferred base being triethylamine to producethe compound of formula XXXIX. Any conventional method to react thiolwith halide can be utilized to carry out the reaction of step (w).

The compound of formula II where Z is

m is 0, r is 1, t is O or I and n is 1 or 2, R⁴ is H, i.e. compounds offormula:

wherein t, n, A and R¹ are as above, can be prepared from the compoundof formula VIII via the reaction scheme in Scheme 15.

In the reaction scheme of Scheme 15, t, n, A and R¹ are as above. R⁶ isan alkyl group having 1 to 7 carbon atoms.

The compound of formula VIII is prepared in same manner as describedhereinbefore in connection with the reaction of step (a) or (b) inScheme 1.

The compound of formula XXVI is prepared from compound of formula VIIIin the same manner as described hereinbefore in connection with thereaction of step (n) in Scheme 5.

The compound of formula XXVI is reacted with compound of formula XL inthe presence of base preferred base being triethylamine to producecompound of formula XLI. Any conventional method to react thiol with1-bromoketone can be utilized to carry out the reaction of step (x).

The compound of formula XLI is the compound of formula II where R¹ is analkyl group containing from 1 to 7 carbon atoms. The compound of formulaXLI can be converted to the free acid i.e. the compound of formula IIwhere R¹ is H by ester hydrolysis. Any conventional method of esterhydrolysis will produce the compound of formula II where R¹ is H.

The compound of formula II where Z is

r is 0, m is 1, t is 0 or 1 and n is 1 or 2, i.e. compounds of formula:

wherein n, t and A are as above, can be prepared from the compound offormula XLII via the reaction scheme in Scheme 16.

In Scheme 16, t, n, and A are as above. Y is a leaving group such ashalide, mesyloxy or tosyloxy. Y¹ is halo preferably bromo.

In Scheme 16, the compound of formula XLII is converted to compound offormula XLIII via the reaction of step (y) by selective displacement ofhydroxyl group of primary alcohol by halogen. Any conventionalhalogenating agent can be utilized to carry out this reaction with thepreferred halogenating agent being phosphorous tribromide. This reactionis carried out in low temperature. Any conditions conventional for thismethod can be utilized to carry out the reaction of step (y). Thecompound of formula XLIII was used immediately without furtherpurification.

The compound of formula XLIII is reacted with compound of formulaXXXVIII in the presence of base to produce the compound of formula XLIV.Any conventional method of condensing thiol with halide can be utilizedto carry out the reaction of step (z). Any conventional base can beutilized to carry out this reaction with the preferred base beingtriethylamine.

The compound of formula XLIV is converted to the compound of formula XLVby reaction with compound of formula VII via the reaction of step (a′).This reaction is carried out in the same manner as describedhereinbefore in connection with reaction of step (a) in Scheme 1.

The compound of formula II where Z is

r is 0, m is 1, t is 0 or 1 and n is 1 or 2, R⁴ is H, i.e. the compoundsof formula:

wherein A and R¹ are as above, can be prepared from the compound offormula XLII via the reaction scheme in Scheme 16.

In Scheme 16, t, n, and A are as above. Y is a leaving group such ashalide, mesyloxy or tosyloxy. Y¹ is halo preferably bromo. R⁶ is analkyl group having 1 to 7 carbon atoms.

The compound of formula XLII is converted to compound of formula XLIIIin the same manner as described hereinbefore in connection with thereaction of step (y).

The compound of formula XLIII is reacted with compound of formula XL viareaction of step (b′) as described in connection with the reaction ofstep (x) in Scheme 15.

The compound of formula XLVI is converted to compound of formula XLVIIvia reaction of step (c′). This reaction is carried out in the samemanner as described in the reaction of step (a) or (b) in Scheme 1.

The compound of formula XLVII is the compound of formula II where R¹ isan alkyl group containing from 1 to 7 carbon atoms. The compound offormula XLVII can be converted to the free acid i.e. the compound offormula II where R¹ is H by ester hydrolysis. Any conventional method ofester hydrolysis will produce the compound of formula II where R¹ is H.

The compound of formula II where Z is

r is 0, m is 1, t is 0 or 1 and n is 1 or 2, i.e. compounds of formula:

wherein t, n, A and R¹ are as above, R⁴ is —NHCO₂C(CH₃)₃, —NHCH₃,—NHCH₂CH₃, can be prepared from the compound of formula XLIII viareaction scheme in Scheme 17.

In Scheme 17, t, n, A and R¹ are as above. Y is a leaving group such ashalide, mesyloxy or tosyloxy. R⁷ is an alkyl group having 1 to 7 carbonatoms and R⁸ is an alkyl group containing from 1 to 2 carbon atoms. Y¹is halo preferably bromo.

In Scheme 17, the compound of formula XLIII (prepared in the same manneras described hereinbefore in reaction of step (y) in Scheme 16) isreacted with the compound of formula XXXII in the presence of a base toproduce the compound of formula XLVIII via reaction of step (d′). Incarrying out this reaction it is generally preferred to utilizetriethylamine as base. Any conventional method of reacting Boc-cyst-OEtwith halide can be utilized to carry out this reaction.

The compound of formula XLIX is produced by reacting compound of formulaXLVIII with compound of formula VII or IX. This reaction is carried outin the same manner as described in the reaction of step (a) or (b) inScheme 1.

The compound of formula XLIX is compound of formula II where R⁴ is—NHCO₂C(CH₃)₃ and R¹ is an alkyl group having 2 carbon atoms.

The compound of formula XLIX can be converted to the free acid i.e. thecompound of formula II where R¹ is H by ester hydrolysis. Anyconventional method of ester hydrolysis will produce the compound offormula II where R¹ is H and R⁴ is —NHCO₂C(CH₃)₃—.

The compound of formula XLIX is converted to compound of formula L firstvia reaction of step (f′) by deprotecting t-butoxy group usingtrifluoroacetic acid and replacing by lower alkyl containing from 1 to 2carbon atoms via reaction of step (g′). Any conventional method tocondense amine with alkyl halide can be used to carry out this reaction.

The compound of formula L is compound of formula II where R⁴ is an aminehaving 1 to 2 carbon atoms and R¹ is an alkyl group having 2 carbonatoms.

The compound of formula L can be converted to the free acid i.e. thecompound of formula LI where R¹ is H by basic hydrolysis via reaction ofstep (h′).

The compound of formula LI is compound of formula II where R⁴ is —NHCH₃or —NHCH₂CH₃ and R¹ is H. Any conventional method of ester hydrolysiswill produce the compound of formula II where R¹ is H.

The compound of formula LI can be converted to compound of formula LIIwhere R⁷ is an alkyl group having 1 to 7 carbon atoms by esterificationof carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (i′).

The compound of formula LII is compound of formula II where R¹ is analkyl group having 1 to 7 carbon atoms and R⁴ is —NHCH₃ or —NHCH₂CH₃—.

The compound of formula III

wherein n is 1 or 2 and A is as above, can be prepared from the compoundof formula LIII, via reaction scheme in Scheme 18, wherein n, A and Yare as above.

In Scheme 18, the compound of formula LIII is converted to the compoundof formula LIV in the same manner as described in connection withreaction of step (a) or (b) in Scheme 1.

The compound of formula LIV is converted to the compound of formula IIIvia reaction of step (k′) by heating the compound of formula LIV withsodium azide in the presence of ammonium chloride in dimethylformamide.Any conventional conditions to convert nitrile to terazole can beutilized to carry out this reaction.

The compound of formula IV

wherein R¹ is as above, can be prepared from 2′,6′-difluoroacetophenonevia reaction scheme in Scheme 19.

In Scheme 19, R⁶ is an alkyl group having 1 to 7 carbon atoms.

The compound of formula LV is converted to compound of formula LVI viareaction of step (l′) in same manner as described hereinbefore inconnection with reaction of step (c) in Scheme 1.

The compound of formula LVI is the compound of formula IV where R¹ is analkyl group having 1 to 7 carbon atoms. The compound of formula LVI canbe converted to the free acid i.e. the compound of formula IV where R¹is H by ester hydrolysis. Any conventional method of ester hydrolysiswill produce the compound of formula IV where R¹ is H.

The compound of formula V

wherein n, A and R¹ are as above, R¹⁴ is hydroxy can be prepared fromthe compound of formula VI, via reaction scheme in Scheme 20.

In Scheme 20, n, A are as above. Y is a leaving group such as halide,mesyloxy or tosyloxy. R⁷ is an alkyl group having 1 to 7 carbon atomsand R⁸ is an alkyl group having from 1 to 2 carbon atoms.

The compound of formula VI is converted to compound of formula VIII insame manner as described hereinbefore in connection with the reaction ofstep (a) or (b) of Scheme 1.

The compound of formula VIII is reacted with compound of formula LVIIvia reaction of step (m′) in the presence of freshly prepared sodiumalkoxide at room temperature to produce compound of formula LVIII. Anyconventional conditions for this alkylation reaction can be utilized tocarry out the reaction.

The compound of formula LVIII is the compound of formula V where R¹ isan alkyl group having 1 to 2 carbon atoms. The compound of formula LVIIIcan be converted to the free acid i.e. the compound of formula V whereR¹ is H by ester hydrolysis via reaction of step (n′). Any conventionalmethod of ester hydrolysis will produce the compound of formula V whereR¹ is H.

The compound of formula LVIII can be converted to compound of formulaLIX where R⁷ is an alkyl group having 1 to 7 carbon atoms byesterification of carboxylic acid with compound of formula XXX usingN,N-dicyclohexylcarbodiimide as dehydrating condensing agent. Anyconditions conventional for this reaction can be utilized to carry outthe reaction of step (o′).

The compound of formula LIX is the compound of formula V where R¹ is analkyl group having 1 to 7 carbon atoms.

The compound of formula I′ where X is —CH₂CH₂—, R⁹ is hydrogen, halo, oralkoxy having 1 to 3 carbon atoms, q and m are 0, t is 0 or 1, and n is1 or 2, i.e. compounds of formula:

wherein Q is NR¹⁰R¹¹ where R¹⁰ is hydrogen and R¹¹ is hydroxyl group. t,n, A, and R⁹ are as described above, can be prepared from the compoundof the formula

via the reaction scheme of Scheme 21.

In the reaction scheme 21, A, t, R⁹, R⁶ and n are as above.

The compound of formula XI is prepared in the same manner as describedin the reaction scheme of Scheme 1.

The compound of formula XI can be converted to the compound of formulaXCII via reaction step (s″) by treating the compound of formula XI withhydroxylamine hydrochloride in an organic solvent, for example ethanol,tetrahydrofuran or the like. The reaction is carried out using organicbase for example, potassium hydroxide or the like. Any conditionsconventional for the synthesis of hydroxamic acids can be utilized tocarry out this reaction.

The compound of formula I′ where X is —CH₂—CH₂—, q and m are 0, t is 0or 1, and n is 1 or 2, R⁹ is hydrogen, halo, or alkoxy having 1 to 3carbon atoms, i.e. compounds of formula:

wherein t, n, A, and R⁹ are described as above. Q is NR¹⁰R¹¹ where R¹⁰and R¹¹ are hydrogen.can be prepared from the compound of the formula

via the reaction scheme in Scheme 21.

In the reaction scheme of Scheme 21, A, t, R⁹ and n are as above. R¹ isH, R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula XI is prepared in the same manner as describedin the reaction scheme of Scheme 1. The compound of formula XI is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XI can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.

The compound of formula XI can be converted to compound of formula XCIIIvia reaction step (t″) by first activating by for example,benzotriazole-1-yloxytrispyrrolidinophosphonium hexafluorophosphate, orthe like in an organic solvent, for example, methylene chloride,N,N-dimethylformamide or the like followed by addition of aqueousammonium hydroxide or ammonia. The reaction is carried out using organicbase for example, triethylamine, diisopropylethylamine or the like.

Any conditions conventional to synthesize amide can be utilized to carryout the reaction of step (t″).

The compound of formula I′ where X is —CH₂—CH₂—, q and m are 0, t is 0or 1, and n is 1 or 2, R⁹ is hydrogen, halo, or alkoxy having 1 to 3carbon atoms, i.e. compounds of formula:

wherein t, n, A, and R⁹ are described as above. Q is NR¹⁰R¹¹ where R¹⁰and R¹¹ are independently hydrogen or alkyl having 1 to 3 carbon atomscan be prepared from the compound of the formula

via the reaction scheme in Scheme 21.

In the reaction scheme of Scheme 21, A, t, R⁹ and n are as above. R¹ isH and R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula XI is prepared in the same manner as describedin the reaction scheme of Scheme 1. The compound of formula XI is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XI can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.

The compound of formula XI can be converted to the compound of formulaXCIV either by first reacting with a chlorinating reagent for example,thionyl chloride or the like then reacting acid halide withcorresponding amine. Any conventional method of condensing amine with anacid halide can be utilized to carry out the reaction of step (u″) or bycondensing corresponding amine with the compound of formula XI using1,3-Dicyclohexylcarbodiimide as condensing agent.

Any conventional method of condensing amine with an acid can be utilizedto carry out the reaction of step (u″).

The compound of formula I′ where X is —CH₂—CH₂—, q is 1, R⁵ is an alkylgroup having 1 to 3 carbon atoms, R⁹ is hydrogen, halo, or alkoxy having1 to 3 carbon atoms, m is 0, t is 0 or 1, and n is 1 or 2, i.e.compounds of formula:

wherein Q is NR¹⁰R¹¹ where R¹⁰ is hydrogen and R¹¹ is hydroxyl group. A,t, n, and R⁹ are as described above, can be prepared from the compoundof the formula

via the reaction scheme in Scheme 22.

In the reaction scheme 22, q, A, t, R⁵, R⁹ and n are described as above.R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula XXV is prepared in the same manner as describedin the reaction scheme of Scheme 4.

The compound of formula XXV can be converted to the compound of formulaXCV via reaction step (v″) in the same manner as described in reactionstep (s″) of Scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, q is 1, R⁵ is an alkylgroup having 1 to 3 carbon atoms, R⁹ is hydrogen, halo, or alkoxy having1 to 3 carbon atoms, m is 0, t is 0 or 1, and n is 1 or 2, R¹ is H, i.e.compounds of formula:

wherein q, t, n, A, R⁵ and R⁹ are described as above. Q is NR¹⁰R¹¹ whereR¹⁰ and R¹¹ are hydrogen and can be prepared from the compound of theformula

via the reaction scheme in Scheme 22.

In the reaction scheme of Scheme 22, q, A, t, R⁵, R⁹ and n are as above.R¹ is H and R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula XXV is prepared in the same manner as describedin the reaction scheme of Scheme 4. The compound of formula XXV is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XXV can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.

The compound of formula XXV can be converted to the compound of formulaXCVI via reaction step (w″) in the same manner as described in step (t″)of reaction scheme 21. The compound of formula I′ where X is —CH₂—CH₂—,q is 1, R⁵ is an alkyl group having 1 to 3 carbon atoms, R⁹ is hydrogen,halo, or alkoxy having 1 to 3 carbon atoms, m is 0, t is 0 or 1, and nis 1 or 2, i.e. compounds of formula:

wherein q, t, n, A, R⁵ and R⁹ are as described above, Q is NR¹⁰R¹¹ whereR¹⁰ and R¹¹ are independently hydrogen or alkyl having 1 to 3 carbonatoms can be prepared from the compound of the formula

via the reaction scheme in Scheme 22.

In the reaction scheme 22, q, A, t, R⁵, R⁹ and n are as above. R⁶ is analkyl having 1 to 7 carbon atoms. R¹ is H.

The compound of formula XXV is prepared in the same manner as describedin the reaction scheme of Scheme 4. The compound of formula XXV is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XXV can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.

The compound of formula XXV can be converted to the compound of formulaXCVII via reaction step (x″) in the same manner as described in step(u″) of reaction scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms, m is 1, q is 0, t is 0 or 1 and nis 1 or 2, i.e. compounds of the formula:

wherein Q is NR¹⁰R¹¹ where R¹⁰ is hydrogen, R¹¹ is hydroxyl group. t, n,A, and R⁹ are as described above, can be prepared from the compound ofthe formula

via the reaction scheme in Scheme 23.

In the reaction scheme 23, A, t, R⁹ and n are as described above. R⁶ isan alkyl having 1 to 7 carbon atoms.

The compound of formula LXXXV is prepared in the same manner asdescribed in the reaction scheme of Scheme 11.

The compound of formula LXXXV can be converted to the compound offormula XCVIII via reaction step (y″) in the same manner as described inreaction step (s″) of Scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms, m is 1, q is 0, t is 0 or 1 and nis 1 or 2, i.e. compounds of the formula:

wherein t, n, A, and R⁹ are described as above. Q is NR¹⁰R¹¹ where R¹⁰and R¹¹ are hydrogen.can be prepared from the compound of the formula

via the reaction scheme in Scheme 23.

In the reaction scheme of Scheme 23, A, t, R⁵, R⁹ and n are as above. R¹is H. R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula LXXXV is prepared in the same manner asdescribed in the reaction scheme of Scheme 11. The compound of formulaLXXXV is the compound of formula I′ where R¹ is an alkyl groupcontaining from 1 to 7 carbon atoms. The compound of formula LXXXV canbe converted to the free acid i.e. the compound of formula I′ where R¹is H by ester hydrolysis.

The compound of formula LXXXV can be converted to the compound offormula XCIX via reaction step (z″) in the same manner as described inreaction step (t″) of reaction scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms, m is 1, q is 0, t is 0 or 1 and nis 1 or 2, i.e. compounds of the formula:

wherein t, n, A, and R⁹ are as described above, Q is NR¹⁰R¹¹ where R¹⁰and R¹¹ are independently hydrogen or alkyl having 1 to 3 carbon atoms.can be prepared from the compound of the formula

via the reaction scheme in Scheme 23.

In the reaction scheme 23, A, t, R⁹ and n are as above. R¹ is H. R⁶ isan alkyl having 1 to 7 carbon atoms.

The compound of formula LXXXV is prepared in the same manner asdescribed in the reaction scheme of Scheme 11. The compound of formulaLXXXV is the compound of formula I′ where R¹ is an alkyl groupcontaining from 1 to 7 carbon atoms. The compound of formula LXXXV canbe converted to the free acid i.e. the compound of formula I′ where R¹is H by ester hydrolysis.

The compound of formula LXXXV can be converted to the compound offormula C via reaction step (a′4″) in the same manner as described instep (u″) of reaction scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms, q is 1, R⁵ is an alkyl grouphaving 1 to 3 carbon atoms, m is 1, t is 0 or 1, and n is 1 or 2, i.e.compounds of formula:

wherein Q is NR¹⁰R¹¹ where R¹⁰ is hydrogen and R¹¹ is hydroxyl group. t,n, A, R⁵ and R⁹ are as described above,can be prepared from the compound of the formula

via the reaction scheme in Scheme 24.

In the reaction scheme 24, q, A, t, n, R⁵, R⁹, and R⁶ are described asabove.

The compound of formula XC is prepared in the same manner as describedin the reaction scheme of Scheme 13.

The compound of formula XC can be converted to the compound of formulaCII via reaction step (b′″) in the same manner as described in reactionstep (s″) of Scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms, q is 1, R⁵ is an alkyl grouphaving 1 to 3 carbon atoms, m is 1, t is 0 or 1, and n is 1 or 2, i.e.compounds of formula:

wherein q, t, n, A, R⁵ and R⁹ are described as above. Q is NR¹⁰R¹¹ whereR¹⁰ and R¹¹ are hydrogen.can be prepared from the compound of the formula

via the reaction scheme in Scheme 24.

In the reaction scheme of Scheme 24, q, A, t, R⁵, R⁹ and n are as above.R¹ is H. R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula XC is prepared in the same manner as describedin the reaction scheme of Scheme 13. The compound of formula XC is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XC can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.

The compound of formula XC can be converted to the compound of formulaCIII via reaction step (c′″) in the same manner as described in step(t″) of reaction scheme 21.

The compound of formula I′ where X is —CH₂—CH₂—, R⁹ is hydrogen, halo,or alkoxy having 1 to 3 carbon atoms, q is 1, R⁵ is an alkyl grouphaving 1 to 3 carbon atoms, m is 1, t is 0 or 1, and n is 1 or 2, i.e.compounds of formula:

wherein q, t, n, A, R⁵ and R⁹ are as described above, Q is NR¹⁰R¹¹ whereR¹⁰ and R¹¹ are independently hydrogen or alkyl having 1 to 3 carbonatoms.can be prepared from the compound of the formula

via the reaction scheme in Scheme 24.

In the reaction scheme 24, q, A, t, R⁵, R⁹ and n are as above. R¹ is H.R⁶ is an alkyl having 1 to 7 carbon atoms.

The compound of formula XC is prepared in the same manner as describedin the reaction scheme of Scheme 13. The compound of formula XC is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XC can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.

The compound of formula XC can be converted to the compound of formulaCIV via reaction step (d′″) in the same manner as described in step (u″)of reaction scheme 21.

The compound of formula V′ where n is 1 or 2, t is 0, R¹, R⁹ and R¹⁴ areH, i.e. compounds of formula:

wherein t, n, A, R⁹, R¹⁴ and R¹ are as described above,can be prepared from the compound of the formula

via the reaction scheme in Scheme 25.

In the reaction scheme of Scheme 25, A, t, and n are as above. R⁶ is analkyl group containing from 1 to 7 carbon atoms.

The compound of formula XI is prepared in the same manner as describedin the reaction scheme of Scheme 1.

The compound of formula XI can be converted to compound of formula CVvia reaction step (e′″), by treating the compound of formula XI withbromine or the like in an organic solvent, for example ether, carbontetrachloride with the preferred organic solvent being ether.

As the reaction temperature, ice cooling to room temperature can be usedwith the preferred being ice cooling.

The compound of formula CV can be converted to the compound of formulaCVI via reaction step (f′″), by dehydrobromination. The reaction iscarried out using conventional base preferred base being triethylamineor the like in an organic solvent for example carbon tetrachloride orthe like. Any of the conditions conventional in dehydrobromination canbe utilized to carry out the reaction of step (f′″).

The compound of formula CVI is the compound of formula V′ where R¹ is analkyl group containing from 1 to 7 carbon atoms. The compound of formulaCVI can be converted to the free acid i.e. the compound of formula V′where R¹ is H by ester hydrolysis. Any conventional method of esterhydrolysis will produce the compound of formula V′ where R¹ is H.

The compound of formula CXVI where X is —CH₂—CH₂—, t is 0 or 1, and n is1 or 2, R¹ and R⁹ are H, i.e. compounds of formula:

wherein t, n, A, and R⁹ are as described above, R¹ is H.can be prepared from the compound of the formula

via the reaction scheme in Scheme 26.

In the reaction scheme of 26, A, t, n and R⁹ are as above. R¹ is H. R⁶is an alkyl group having 1 to 7 carbon atoms.

The compound of formula XI is prepared in the same manner as describedin the reaction scheme of Scheme 1. The compound of formula XI is thecompound of formula I′ where R¹ is an alkyl group containing from 1 to 7carbon atoms. The compound of formula XI can be converted to the freeacid i.e. the compound of formula I′ where R¹ is H by ester hydrolysis.Any conventional method of ester hydrolysis will produce the compound offormula I′ where R¹ is H.

The compound of formula XI is converted to compound of formula CVII viareaction step (g′″) via Wolff-Kishner reduction by treating the compoundof formula XI with hydrazine hydrate and potassium hydroxide in anorganic solvent for example, ethylene glycol or the like. Any of theconditions conventional in Wolff-Kishner reductions can be utilized tocarry out the reaction of step (g′″).

The compound of formula XCI where n is 1 or 2, R⁹ is H and R¹ ishydrogen or alkyl having 1 to 3 carbon atoms, i.e. compounds of formula:

wherein n, A, R⁹ and R¹ are as described above,can be prepared from the compound of the formula

via the reaction scheme in Scheme 27.

In the reaction scheme of Scheme 27, R⁹ is a hydrogen atom, t is 0, R⁶is an alkyl having 1 to 7 carbon atoms, A and n are as described above.

The compound of formula XI is prepared in the same manner as describedin the reaction scheme of Scheme 1.

The compound of formula XI can be converted to the compound of formulaCVIII via reaction step (h′″) by selectively reducing ketone group to analcohol. This reaction is carried out utilizing conventional reducingagents for example, sodium borohydride in ethanol,Bis-3-methyl-2-butyl-borane in tetahydrofuran or the like. Any of theconditions conventional in such selective reduction reactions can beutilized to carry out the reaction of step (h′″).

The compound of formula CVIII can be converted to compound of formulaCIX via reaction step (i′″) by bromination of compound of formula CVIIIwith brominating reagents for example, phosphorous tribromide intetrahydrofuran or dioxane, hydrogen bromide in acetic acid or dioxane,carbon tetrabromide and bis-(1,2-diphenylphosphino)ethane or the like.Any of the conditions conventional in such bromination reactions can beutilized to carry out the reaction of step (i′″).

The compound of formula CIX can be converted to the compound of formulaCX via reaction step (j′″), by dehydrobromination. The reaction iscarried out using conventional base preferred base being triethylamineor the like in an organic solvent for example carbon tetrachloride orthe like. Any of the conditions conventional in such dehydrobrominationreactions can be utilized to carry out the reaction of step (j′″). Thecompound of formula CX is the compound of formula XCI where R¹ is analkyl group containing from 1 to 3 carbon atoms. The compound of formulaCX can be converted to the free acid i.e. the compound of formula XCIwhere R¹ is H by ester hydrolysis. Any conventional method of esterhydrolysis will produce the compound of formula XCI where R¹ is H.

The compound of formula CXVII where X is —CH₂—CH₂—, and n is 0 or 2, R¹⁵is a hydrogen or lower alkyl group having 1 to 3 carbon atoms, R⁹ ishydroxy, hydrogen, alkoxy group having 1 to 3 carbon atoms, halogenatom, R¹ is hydrogen or alkyl having 1 to 3 carbon atoms, i.e. compoundsof formula:

wherein n, A, R⁹ and R¹⁵ are as described above,can be prepared by reacting the compound of the formula

with compound of formula

via the reaction scheme in Scheme 28.

In the reaction scheme of Scheme 28, A, n, R⁹, R¹⁵ are described asabove, R⁶ is an alkyl having 1 to 3 carbon atoms.

The compound of formula CXI can be converted to compound of formulaCXIII via reaction step (k′″) by treating compound of formula CXI withcondensing agent, for example diethyl cyanophosphate,1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide or the like in an organicsolvent, for example, methylene chloride, N,N-dimethylformamide followedby addition of compound of formula CXII.

The reaction temperature can be from 0° C. to room temperature.

The compound of formula CXIII can be converted to the compound offormula CXIV via reaction of step (l′″) by alkylating the compound offormula CXIII with the compound of formula X. This reaction is carriedout in the same manner as described in the reaction step (c) of reactionscheme 1.

The compound of formula CXIV is the compound of formula CXVII where R⁹is an alkoxy group having 1 to 3 carbon atoms, halogen atom. The R⁹ canbe converted to hydroxy via demethylation by using for example, borontribromide in methylene chloride or the like. Any of the conditionsconventional in such demethylation reactions can be utilized to carryout the reaction.

The compound of formula CXIV is the compound of formula CXVII where R¹is an alkyl group having 1 to 3 carbon atoms. The compound of formulaCXIV can be converted to the free acid i.e. the compound of formulaCXVII where R¹ is H by ester hydrolysis. Any conventional method ofester hydrolysis will produce the compound of formula CXVII where R¹ isH.

Compounds of general formula CXI can be prepared by etherification ofcompound of formula CI by using alkyl halide followed by an esterhydrolysis.

wherein R¹⁶ is a lower alkyl group having 1 to 3 carbon atoms. R⁹ is ahydroxyl group.

The reaction between compound of formula CI and alkyl halide can becarried like in an organic solvent, for example N,N-dimethylformamide orthe like, using base, for example potassium carbonate, cesium carbonateor the like. Any of the conditions conventional in such alkylationreactions can be utilized to carry out this reaction. Ester hydrolysiscan be conducted under acidic conditions for example, hydrochloric acidor hydrochloric acid mixed with organic solvent for example, ethanol orusing acetic acid or the like. The reaction can be carried out at roomtemperature to solvent refluxing temperature. Any conventionalconditions for acidic ester hydrolysis can be utilized to carry out thisreaction. Further, if needed, ester hydrolysis can be carried out usingbasic conditions, for example, in an aqueous solution of sodiumhydroxide or a mixed solution of sodium hydroxide in an organic solventfor example, ethanol or the like. Any conditions conventional for basichydrolysis can be utilized to carry out this reaction

Compounds of general formula CXII can be prepared by reacting compoundof formula VII with chlorinating agent for example, trimethylsilylchloride, thionyl chloride or the like in an organic solvent forexample, dimethyl sulfoxide, N,N-dimethylformamide or the like. Thereaction temperature can be room temperature to organic refluxingtemperature. Any conditions conventional for chlorination reactions canbe utilized to carry out the reaction.

The chloromethyl intermediate was converted to compound of formula CXIIvia Gabriel synthesis by treating chloromethyl intermediate withpotassium phthalimide in an organic solvent for example,N,N-dimethylformamide, dioxane or the like. The phthalimide is thenreacted with hydrazine by an exchange reaction in an organic solvent forexample, ethanol, dioxane or the like to produce the compound of formulaCXII. Any of the conditions conventionally used in the Gabriel synthesiscan be utilized to carry out the reaction.

The invention will be better understood by reference to the followingexamples which illustrate but do not limit the invention describedherein.

CHEMICAL SYNTHESIS EXAMPLES Example 1 Synthesis of4-(4-(2-Fluorobenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(2-Fluorobenzyloxy)acetophenone

A solution of 4-Hydroxyacetophenone (2.80 g, 20.6 mmol) in dry DMF (15ml) was added at room temperature to a suspension of NaH (60% in oil,0.794 g) in dry DMF (20 ml). When evolution of hydrogen ceased,2-Fluorobenzyl bromide (3 g, 15.8 mmol) was added drop wise. Thereaction mixture was stirred at room temperature for 6 hours, quenchedwith sat aq. NH₄Cl and concentrated in vacuo. The crude residue wastaken in EtOAc and washed with water and brine. The organic layer wasdried over Na₂SO₄, filtered and concentrated. The residue was purifiedby flash chromatography on silica gel column (hex:ethyl acetate, 2:1) toprovide the title compound as an off white solid.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.1 (m, 4H);7.2-7.3 (m, 1H); 7.4 (t, 1H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(2-fluorobenzyloxy)phenyl)-4-oxobutyrate

To a stirred solution of 4-(2-fluorobenzyloxy)acetophenone (Step A, 1.5g, 6.1 mmol) in dry THF (20 ml) and DMPU (5 ml) was added a solution oflithium bis(trimethylsilyl)amide (1.0M, 7 ml) at −60° C. under argon.After 10 minutes of stirring at −60° C., tert-Butyl bromoacetate (4.75g, 24.4 mmol) was added rapidly. The reaction mixture was stirred for anadditional 10 minutes and then warmed to room temperature for 4 hours.The crude mixture was taken in EtOAc and washed with water and brine.The aqueous layer was extracted one more time with EtOAc. The combinedorganic layers were dried over Na₂SO₄, filtered, concentrated andpurified by flash chromatography on a silica gel column (hex:ethylacetate, 2:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.7 (t, 2H); 3.2 (t, 2H); 5.1 (s,2H); 6.9-7.1 (m, 4H); 7.2-7.3 (m, 1H); 7.4 (t, 1H); 7.9 (d, 2H).

Step C: Preparation of 4-(4-(2-Fluorobenzyloxy)phenyl)-4-oxobutyric acid

A solution of tert-Butyl 4-(4-(2-fluorobenzyloxy)phenyl)-4-oxobutyrate(Step B, 1.27 g, 4.2 mmol) in dichloromethane (25 ml) was treated withtrifluoroacetic acid (5 ml). The reaction mixture was stirred at ambienttemperature for 3 hours and concentrated in vacuo. The purification wasdone by flash chromatography on silica gel column (chloroform:methanol,95:5 spiked with acetic acid) to afford the title compound as a whitepowder.

¹H NMR (270 MHz, CDCl₃:CD₃OD): 2.6 (t, 2H); 3.2 (t, 2H); 5.1 (s, 2H);6.9-7.1 (m, 4H); 7.2-7.3 (m, 1H); 7.4 (t, 2H); 7.9 (d, 2H).

Example 2 Synthesis of 4-(4-(2-Methoxybenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2-Methoxybenzyloxy)acetophenone

A solution of 2-Methoxybenzyl alcohol (2.99 g, 21.7 mmol) in dry THF (5ml) and dry DMF (5 ml) was added to a stirred solution of4-Hydroxyacetophenone (3.25 g, 23.8 mmol), triphenylphosphine (7.36 g,28.0 mmol), and diethyl azodicarboxylate (4.51 g, 25.9 mmol) in dry THF(20 ml) at 5-10° C. The reaction mixture was stirred at O° C. for 2hours, warmed to room temperature and concentrated in vacuo. The residuewas taken in EtOAc and washed twice with saturated NaHCO₃. The organiclayer was dried over Na₂SO₄, filtered, concentrated and purified byflash chromatography on a silica gel column (choloroform:methanol, 99:1)to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 3.9 (s, 3H); 5.2 (s, 2H); 6.9-7.1(m, 4H); 7.3 (m, 1H); 7.4 (d, 1H); 7.9 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2-methoxybenzyloxy)phenyl)-4-oxobutyrate

To a stirred solution of 4-(2-Methoxybenzyloxy)acetophenone (Step A,1.22 g, 4.7 mmol) in dry THF (20 ml) and DMPU (5 ml) was added asolution of lithium bis(trimethylsilyl)amide (1.0M, 5 ml) under argon at−60° C. After 10 minutes of stirring at −60° C., ethyl bromoacetate(2.59 g, 15.6 mmol) was added rapidly. The reaction mixture was stirredfor an additional 10 minutes and then warmed to room temperature for 2hours. The crude mixture was taken in EtOAc and washed with water. Theaqueous layer was extracted one more time with EtOAc and combinedorganic layers were dried over Na₂SO₄, filtered and concentrated. Thepurification was done by flash chromatography on silica gel column(hex:ethyl acetate, 4:1) to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.6 (t, 2H); 3.2 (t, 2H); 3.8 (s,3H); 4.1 (q, 2H); 5.1 (s, 2H); 6.9-7.0 (m, 4H); 7.1-7.3 (m, 2H); 7.9 (d,2H).

Step C: Preparation of 4-(4-(2-Methoxybenzyloxy)phenyl)-4-oxobutyricacid

A solution of Ethyl 4-(4-(2-methoxybenzyloxy)phenyl)-4-oxobutyrate (StepB, 1.49 g, 4.3 mmol) in abs ethanol (20 ml) was treated with 1N NaOH (6ml). The reaction mixture was stirred at room temperature for 2 hoursand then acidified with 1M HCl. The resulting white solid was filtered,washed with cold water and dried under vacuum to provide the titlecompound.

¹H NMR (270 MHz, CDCl₃: CD₃OD): 2.6 (t, 2H); 3.2 (t, 2H); 3.8 (s, 3H);5.1 (s, 2H); 6.9-7.0 (m, 4H); 7.2-7.3 (m, 2H); 7.8 (d, 2H).

Example 3 Synthesis of3-[(4-(2-Fluorobenzyloxy)phenyl)-methylthio]propionic acid

Step A: Preparation of 4-Hydroxybenzyl bromide

To a stirred solution of PBr₃ (1.38 g, 5.0 mmol) in dry THF (2 ml) at−5° C. was added a solution of dry pyridine (0.201 ml) in dry THF (0.4ml). A solution of 4-Hydroxybenzyl alcohol (1.89 g, 15.2 mmol) in dryTHF (23 ml) was added drop wise to the reaction mixture. The reactionmixture was allowed to stand at room temperature for 18 hours, thendiluted with THF and filtered through celite pad. The filtrate wasevaporated, the resulting semisolid was redissolved in dry toluene (16ml). The solution was maintained at −20° C. for 2 hours, and thenfiltered through celite pad to provide the title compound as a lightyellow solution which was used without further purification.

Step B: Preparation of Ethyl 3-((4-hydroxyphenyl)-methylthio)propionate

To a solution of NaH (60% dispersed in oil, 0.731 g, 21.7 mmol) in dryDMF (15 ml) was added Ethyl 3-mercaptopropionate (2.66 g, 19.8 mmol).When the evolution of hydrogen ceased, 4-Hydroxybenzyl bromide from StepA was added. The reaction mixture was stirred for 16 hours at roomtemperature, quenched with sat. NH₄Cl and concentrated in vacuo. Thecrude residue was taken in EtOAc and washed with water and brine. Theaqueous layer was washed one more time with EtOAc. The combined organiclayers were dried over Na₂SO₄, filtered and concentrated. Thepurification was done by flash chromatography on silica gel column(dichloromethane:ethyl acetate, 95:5) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.4-2.6 (m, 4H); 3.6 (s, 2H); 4.1(q, 2H); 6.7 (d, 2H); 7.2 (d, 2H).

Step C: Preparation of Ethyl3-((4-(2-fluorobenzyloxy)phenyl)-methylthio)propionate

To a solution of NaH (60% dispersed in oil, 0.054 g, 1.3 mmol) in dryDMF (10 ml) was added Ethyl 3-((4-hydroxyphenyl)-methylthio)propionate(Step B, 2.5 g, 1.0 mmol). When the evolution of hydrogen ceased,2-Fluorobenzyl bromide (0.263 g, 1.3 mmol) was added. The reactionmixture was stirred for 4 hours at room temperature, quenched with sat.NH₄Cl, and concentrated in vacuo. The crude residue was taken in EtOAcand washed twice with water and brine. The aqueous layer was washed onemore time with EtOAc. The combined organic layers were dried overNa₂SO₄, filtered and concentrated. The purification was done by flashchromatography on silica gel column (hex:ethyl acetate, 4:1) to providethe title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.4-2.6 (m, 4H); 3.6 (s, 2H); 4.2(q, 2H); 5.15 (s, 2H); 6.9 (d, 2H); 7.2-7.4 (m, 5H); 7.5 (t, 1H).

Step D: Preparation of3-((4-(2-fluorobenzyloxy)phenyl)-methylthio)propionic acid

To a solution of Ethyl3-((4-(2-fluorobenzyloxy)phenyl)-methylthio)propionate (Step C, 0.122 g,0.35 mmol) in ethanol (5 ml) was added 1N NaOH (0.5 ml) at roomtemperature. The reaction mixture was stirred for 3 hours, acidifiedwith 1M HCl and concentrated in vacuo to give white solid which waspurified by flash chromatography on silica gel column(chloroform:methanol, 92.5:7.5 spiked with acetic acid) to provide thetitle compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.4-2.6 (m, 4H); 3.7 (s, 2H); 5.1 (s, 2H); 6.9(d, 2H); 7.2-7.4 (m, 5H); 7.5 (t, 1H).

Example 4 Synthesis of 4-(4-(3-Fluorobenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(3-Fluorobenzyloxy)acetophenone

Using the method of Example 1, Step A, using 3-Fluorobenzyl bromide asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.1 (s, 2H); 7.0 (m, 3H); 7.2-7.3(t, 2H); 7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(3-fluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 1, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.7 (t, 2H); 3.2 (t, 2H); 5.1 (s,2H); 7.0 (m, 3H); 7.2 (t, 2H); 7.4 (m, 1H); 8.0 (d, 2H).

Step C: Preparation of 4-(4-(3-Fluorobenzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 1, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s, 2H); 6.9-7.1(m, 3H); 7.2-7.3 (m, 2H); 7.4 (q, 1H); 7.9 (d, 2H).

Example 5 Synthesis of 4-(4-(4-Fluorobenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(4-Fluorobenzyloxy)acetophenone

Using the method of Example 1, Step A, using 4-Fluorobenzyl bromide asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.1 (s, 2H); 7.0 (d, 2H); 7.1 (t,2H); 7.4 (m, 2H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(4-fluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 1, Step B, the title compound was obtained

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s,2H); 7.0 (m, 2H); 7.2 (t, 2H); 7.4 (m, 2H); 8.0 (d, 2H).

Step C: Preparation of 4-(4-(4-Fluorobenzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 1, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s, 2H); 6.9-7.1(m, 2H); 7.2-7.3 (d, 2H); 7.4 (m, 2H); 7.9 (d, 2H).

Example 6 Synthesis of 4-(4-((2-Pyridinyl)-methoxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-((2-Pyridinyl)-methoxy)acetophenone

A solution of 4-Hydroxyacetophenone (1.99 g, 14.6 mmol) in dry DMF (5ml) was added at room temperature to a suspension of NaH (60% in oil,0.604 g) in dry DMF (20 ml). When evolution of hydrogen ceased,2-Picolyl chloride hydrochloride (2 g, 12.1 mmol) was added. Thereaction mixture was stirred at room temperature for 16 hours, quenchedwith sat aq. NH₄Cl and concentrated in vacuo. The crude residue wastaken in EtOAc and washed with water and brine. The aqueous layer waswashed twice with EtOAc. The combined organic layers were dried overNa₂SO₄, filtered and concentrated. The residue was purified by flashchromatography on silica gel column (hex:ethyl acetate, 1:1) to providethe title compound.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 7.0 (d, 2H); 7.2 (m,1H); 7.5 (d, 1H); 7.7 (t, 1H); 7.9 (d, 2H); 8.6 (s, 1H).

Step B: Preparation of tert-Butyl4-(4-((2-pyridinyl)-methoxy)phenyl)-4-oxobutyrate

To a stirred solution of 4-((2-Pyridinyl)-methoxy)acetophenone (Step A,0.968 g, 3.6 mmol) in dry THF (16 ml) and DMPU (4 ml) was added asolution of lithium bis(trimethylsilyl)amide (1.0M, 5 ml) at −60° C.under argon. After stirring for 10 minutes at −60° C., tert-Butylbromoacetate (2.64 g, 13.5 mmol) was added rapidly. The reaction mixturewas stirred for an additional 10 minutes and then warmed to roomtemperature for 4 hours. The crude mixture was taken in EtOAc and washedwith water and brine. The aqueous layer was extracted one more time withEtOAc. The combined organic layers were dried over Na₂SO₄, filtered,concentrated and purified by flash chromatography on silica gel column(hex:ethyl acetate, 2:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.7 (t, 2H); 3.2 (t, 2H); 5.3 (s,2H); 7.0 (d, 2H); 7.2 (m, 1H); 7.5 (d, 1H); 7.7 (t, 1H); 7.9 (d, 2H);8.6 (s, 1H).

Step C: Preparation of 4-(4-((2-Pyridinyl)-methoxy)phenyl)-4-oxobutyricacid

A solution of tert-Butyl4-(4-((2-pyridinyl)-methoxy)phenyl)-4-oxobutyrate (Step C, 1.27 g, 4.2mmol) in dichloromethane (25 ml) was treated with trifluoroacetic acid(5 ml). The mixture was stirred at ambient temperature for 3 hours andconcentrated in vacuo. The purification was done by flash chromatographyon silica gel column (chloroform:methanol, 95:5 spiked with acetic acid)to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃:CD₃OD): 2.7 (t, 2H); 3.2 (t, 2H); 5.3 (s, 2H);7.0 (d, 2H); 7.3 (m, 1H); 7.5 (d, 1H); 7.9 (m, 3H); 8.6 (s, 1H).

Example 7 Synthesis of 4-(4-(Benzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(Benzyloxy)acetophenone

Using the method of Example 1, Step A, using Benzyl bromide as thestarting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.1 (s, 2H); 7.0 (d, 2H); 7.3-7.5(m, 5H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl 4-(4-(benzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 1, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.6 (t, 2H); 3.2 (t, 2H); 5.2 (s,2H); 7.0 (d, 2H); 7.3-7.5 (m, 5H); 7.9 (d, 2H).

Step C: Preparation of 4-(4-(Benzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 1, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s, 2H); 7.0 (d,2H); 7.3-7.5 (m, 5H); 7.9 (d, 2H).

Example 8 Synthesis of 4-(4-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2,6-Difluorobenzyloxy)acetophenone

A solution of 4-Hydroxyacetophenone (3.61 g, 26.5 mmol) in dry DMF (5ml) was added at room temperature to a suspension of NaH (60% in oil,1.21 g) in dry DMF (40 ml). When evolution of hydrogen ceased,2,6-Difluorobenzyl bromide (5 g, 24.1 mmol) was added drop wise. Thereaction mixture was stirred at room temperature for 6 hours, quenchedwith sat aq. NH₄Cl and concentrated in vacuo. The crude residue wastaken in EtOAc and washed with water and brine. The aqueous layer wasextracted one more time with EtOAc. The combined organic layers weredried over Na₂SO₄, filtered and concentrated. The residue was purifiedby flash chromatography on silica gel column (hex:ethyl acetate, 2:1) toprovide the title compound.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 4H);7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyrate

To a stirred solution of 4-(2,6-Difluorobenzyloxy)acetophenone (Step A,0.6 g, 22.8 mmol) in dry THF (60 ml) and DMPU (12 ml) was added asolution of lithium bis(trimethylsilyl)amide (1.0M, 30 ml) at −60° C.under argon. After stirring for 10 minutes at −60° C., tert-Butylbromoacetate (8.97 g, 46 mmol) was added rapidly. The reaction mixturewas stirred for an additional 10 minutes and then warmed to roomtemperature for 4 hours. The crude mixture was taken in EtOAc and washedwith water and brine. The aqueous layer was extracted one more time withEtOAc. The combined organic layers were dried over Na₂SO₄, filtered,concentrated and purified by flash chromatography on a silica gel column(hex:ethyl acetate, 2:1) to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.6 (t, 2H); 3.2 (t, 2H); 5.2 (s,2H); 6.9-7.0 (m, 4H); 7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step C: Preparation of 4-(4-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

A solution of tert-Butyl4-(4-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyrate (Step B, 4.76 g, 12.6mmol) in dichloromethane (40 ml) was treated with trifluoroacetic acid(20 ml). The mixture was stirred at ambient temperature for 3 hours andconcentrated in vacuo. The purification was done by flash chromatographyon silica gel column (chloroform:methanol, 95:5 spiked with acetic acid)to provide the title compound as a white powder.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.2 (s, 2H); 6.9-7.0(m, 4H); 7.4 (m, 1H); 7.9 (d, 2H).

Example 9 Synthesis of 4-(4-(2-Chlorobenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(2-Chlorobenzyloxy)acetophenone

Using the method of Example 1, Step A, using 2-Chlorobenzyl bromide asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 7.0 (d, 2H); 7.2-7.5(m, 4H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(2-chlorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 1, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.6 (t, 2H); 3.2 (t, 2H); 5.2 (s,2H); 7.0 (d, 2H); 7.2-7.5 (m, 4H); 7.9 (d, 2H).

Step C: Preparation of 4-(4-(2-Chlorobenzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 1, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃:CD₃OD): 2.6 (t, 2H); 3.2 (t, 2H); 5.2 (s, 2H);7.0 (d, 2H); 7.2 (m, 2H); 7.3 (m, 1H); 7.4 (m, 1H); 7.9 (d, 2H).

Example 10 Synthesis of4-(4-(2-(2-Fluorophenyl)ethoxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(2-(2-fluorophenyl)ethoxy)acetophenone

Using the method of Example 2, Step A, using 2-Fluorophenethyl alcoholas the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 3H); 2.9 (t, 2H); 4.2 (t, 2H); 6.9 (d,2H); 7.1 (m, 2H); 7.3 (m, 2H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(2-(2-fluorophenyl)ethoxy)phenyl)-4-oxobutyrate

Using the method of Example 2, Step B, using tert-Butyl bromoacetate asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.6 (t, 2H); 3.2 (m, 4H); 4.2 (t,2H); 6.9 (d, 2H); 7.1 (m, 2H); 7.3 (t, 2H); 7.9 (d, 2H).

Step C: Preparation of4-(4-(2-(2-Fluorophenyl)ethoxy)phenyl)-4-oxobutyric acid

A solution of tert-Butyl4-(4-(2-(2-Fluorophenyl)ethoxy)phenyl)-4-oxobutyrate (Step 2, 1.2 g, 3.2mmol) in dichloromethane (25 ml) was treated with trifluoroacetic acid(10 ml). The reaction mixture was stirred at ambient temperature for 4hours and concentrated in vacuo. The purification was done by flashchromatography on silica gel column (chloroform:methanol, 95:5 spikedwith acetic acid) to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.3 (t, 2H); 4.2 (t, 2H); 6.9 (d,2H); 7.1 (m, 2H); 7.3 (t, 2H); 7.9 (d, 2H).

Example 11 Synthesis of Ethyl4-(4-(2-Fluorobenzyloxyl)phenyl)-4-oxobutyrate

Step A: Preparation of 4-(4-(2-Fluorobenzyloxy)acetophenone

Using the method of Example 1, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.1 (m, 4H);7.2-7.3 (m, 1H); 7.4 (t, 1H); 7.9 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2-fluorobenzyloxy)phenyl)-4-oxobutyrate

To a stirred solution of 4-(2-Fluorobenzyloxy)acetophenone (7.26 g, 29.7mmol) in dry THF (80 ml) and DMPU (16 ml) was added a solution oflithium bis(trimethylsilyl)amide (1.0M, 35 ml) at −60° C. under argon.After stirring for 10 minutes at −60° C., Ethyl bromoacetate (10.12 g,60.5 mmol) was added rapidly. The reaction mixture was stirred for anadditional 10 minutes and then warmed to room temperature for 4 hours.The crude mixture was taken in EtOAc and washed with water and brine.The aqueous layer was extracted one more time with EtOAc. The combinedorganic layers were dried over Na₂SO₄, filtered, concentrated andpurified by flash chromatography on a silica gel column (hex:ethylacetate, 4:1) to provide the title compound as a white powder.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.7 (t, 2H); 3.2 (t, 2H); 4.2 (q,2H); 5.1 (s, 2H); 6.9 (d, 2H); 7.2 (m, 2H); 7.4 (m, 1H); 7.5 (m, 1H);7.9 (d, 2H).

Example 12 Synthesis of 4-(4-(2-Methylbenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2-Methylbenzyloxy)acetophenone

Using the method of Example 1, Step A, using 2-Methylbenzyl bromide asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 2.5 (s, 3H); 5.2 (s, 2H); 6.9 (d,2H); 7.2-7.3 (m, 3H); 7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(2-methylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 1, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.5 (s, 9H); 2.4 (s, 3H); 2.6 (t, 2H); 3.2 (t,2H); 5.2 (s, 2H); 6.9 (d, 2H); 7.2-7.3 (m, 3H); 7.4 (m, 1H); 7.9 (d,2H).

Step C: Preparation of 4-(4-(2-Methylbenzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 1, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s,2H); 6.9 (d, 2H); 7.2-7.3 (m, 3H); 7.4 (m, 1H); 7.9 (d, 2H).

Example 13 Synthesis of4-[4-(2-(N-(2-fluorobenzyl)-N-methylamino)ethoxy)phenyl]-4-oxobutyricacid

Step A: Preparation of 2-Fluorobenzyl methanesulfonate

To a solution of 2-Fluorobenzyl alcohol (10 g, 79.28 mmol) in drydichloromethane (200 ml) was added triethylamine (12.03 g, 118.9 mmol)under argon at room temperature. Methanesulfonyl chloride (10.71 g, 93.5mmol) was added to the above reaction mixture at 0° C., and stirring wascontinued for another 3 hours. Water (100 ml) was added to the reactionmixture and the mixture was extracted twice with dichloromethane. Thecombined organic layers were washed with water and brine. The reactionmixture was dried over Na₂SO₄, filtered and concentrated to give thetitle compound as an yellow oil which was used without furtherpurification.

¹H NMR (270 MHz, CDCl₃): 1.3 (t, 3H); 2.4-2.6 (m, 4H); 5.25 (s, 2H);6.9-7.5 (m, 4H).

Step B: Preparation of 2-(N-(2-fluorobenzyl)-N-methylamino)-ethanol

A mix of 2-Fluorobenzyl methanesulfonate (Step A, 5 g, 24.5 mmol) and2-(Methylamino)-ethanol (18.4 g, 244.9 mmol) was heated under argon at120° C. with stirring for 7 hours. The mixture was cooled to roomtemperature and concentrated. The crude residue was purified by flashchromatography on silica gel column (chloroform:methanol, 90:10 spikedwith triethylamine) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 3H); 2.6 (m, 2H); 3.6 (m, 4H); 6.9-7.5(m, 4H).

Step C: Preparation of 2-(N-(2-fluorobenzyl)-N-methylamino)-ethylchloride

To a solution of 2-(N-(2-fluorobenzyl)-N-methylamino)-ethanol (Step B,7.51 g, 41 mmol) in dry toluene (50 ml) was added thionyl chloride (16ml). The reaction mixture was stirred at room temperature for 16 hoursand concentrated. The crude mixture was diluted with chloroform andwashed with aq NaHCO₃, water and brine. The organic layer was dried overNa₂SO₄, filtered and concentrated to provide the title compound whichwas used without further purification.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 3H); 2.8 (t, 2H); 3.6 (t, 2H); 3.7 (s,2H); 7.0-7.15 (m, 2H); 7.25 (m, 1H), 7.4 (t, 1H).

Step D: Preparation of4-(2-(N-(2-fluorobenzyl)-N-methylamino)ethoxy)acetophenone

To a solution of 2-(N-(2-fluorobenzyl)-N-methylamino)-ethyl chloride(Step C, 7.48 g, 37 mmol) and 4-Hydroxyacetophenone (10.07 g, 74 mmol)in dry DMF (10 ml) was added K₂CO₃ (7.77 g, 56.2 mmol). The mixture washeated at 80° C. for 6 hours, cooled, quenched with water and extractedtwice with EtOAc. The combined organic layers were dried over Na₂SO₄,filtered and concentrated. The crude residue was purified by flashchromatography on silica gel column (hex:ethyl acetate, 2:1) to providethe title compound as a light yellow oil.

¹H NMR (270 MHz, CDCl₃): 2.35 (s, 3H); 2.4 (s, 3H); 2.8 (t, 2H); 3.7 (s,2H); 4.2 (t, 2H); 6.9 (d, 2H); 7.0-7.15 (m, 2H); 7.25 (m, 1H), 7.4 (t,1H); 7.9 (d, 2H).

Step E: Preparation of tert-Butyl4-[4-(2-(N-(2-fluorobenzyl)-N-methylamino)ethoxy)phenyl]-4-oxobutyrate

Lithium bis(trimethylsilyl)amide (1.0M, 20 ml) was added slowly over 10minutes to a stirred solution of4-(2-(N-(2-fluorobenzyl)-N-methylamino)ethoxy)acetophenone (Step D, 4.91g, 16.3 mmol) in dry THF (60 ml) and DMPU (15 ml) at −65° C. underargon. After 15 minutes of stirring, tert-Butyl bromoacetate (6.35 g,32.6 mmol) was added rapidly. The stirring continued for an additional10 minutes at −65° C. and then reaction was warmed to room temperaturefor 2 hours, quenched with water and extracted twice with EtOAc. Thecombined organic layers were purified by flash chromatography on silicagel column (hex:ethyl acetate, 1:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.5 (s, 9H); 2.4 (s, 3H); 2.6 (t, 2H); 2.8 (t,2H); 3.2 (t, 2H) 3.7 (br, 2H); 4.2 (br, 2H); 6.9 (d, 2H); 7.0-7.15 (m,2H); 7.25 (m, 1H), 7.4 (t, 1H); 7.9 (d, 2H).

Step F: Preparation of4-[4-(2-(N-(2-fluorobenzyl)-N-methylamino)ethoxy)phenyl]-4-oxobutyricacid

A solution of tert-Butyl4-[4-(2-(N-(2-fluorobenzyl)-N-methylamine)ethoxy)phenyl]-4-oxobutyrate(Step E, 2.23 g, 5.3 mol) in dichloromethane (20 ml) was treated withtrifluoroacetic acid (10 ml). The reaction mixture was stirred atambient temperature for 2 hours, and concentrated in vacuo. Thepurification was done by flash chromatography on silica gel column(chloroform:methanol, 92.5:7.5-90:10 spiked with acetic acid) to providethe title compound.

¹H NMR (270 MHz, CDCl₃:CD₃OD): 2.5 (t, 2H); 2.6 (s, 3H); 3.0 (t, 2H);3.4 (t, 2H); 4.2-4.5 (m, 4H); 6.9 (d, 2H); 7.0-7.15 (m, 2H); 7.3 (m,1H), 7.5 (t, 1H), 7.9 (d, 2H).

Example 14 Synthesis of 4-(3-(2-Methylbenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 3-(2-Methylbenzyloxy)acetophenone

Using the method of Example 12, Step A, using 3-Hydroxyacetophenone asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 3H); 2.5 (s, 3H); 5.1 (s, 2H); 7.2-7.3(m, 4H); 7.4 (m, 2H); 7.6 (m, 2H).

Step B: Preparation of tert-Butyl4-(3-(2-methylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 1, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.5 (s, 9H); 2.4 (s, 3H); 2.6 (t, 2H); 3.2 (t,2H); 5.2 (s, 2H); 7.2-7.3 (m, 4H); 7.4 (m, 2H); 7.6 (m, 2H).

Step C: Preparation of 4-(3-(2-Methylbenzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 1, Step C, the title compound was obtained

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.2-7.3 (m, 4H); 7.4 (m, 2H); 7.6 (m, 2H).

Example 15 Synthesis of Ethyl4-(3-(2-fluorobenzyloxyl)phenyl)-4-oxobutyrate

Step A: Preparation of 3-(2-Fluorobenzyloxy)acetophenone

Using the method of Example 1, Step A, using 3-Hydroxyacetophenone asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 7.1 (m, 4H); 7.3 (m,2H); 7.6 (m, 2H).

Step B: Preparation of Ethyl4-(3-(2-fluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 11, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.3 (s, 9H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.1 (t, 2H); 7.2 (d, 2H); 7.4 (m, 1H); 7.5 (t, 1H); 7.6 (d, 2H).

Example 16 Synthesis of Ethyl4-(4-(2-methylbenzyloxyl)phenyl)-4-oxobutyrate

Step A: Preparation of 4-(2-Methylbenzyloxy)acetophenone

Using the method of Example 12, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 2.5 (s, 3H); 5.2 (s, 2H); 6.9 (d,2H); 7.2-7.3 (m, 3H); 7.4 (m, 1H); 8.0 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2-methylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 11, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.4 (s, 3H); 2.7 (t, 2H); 3.2 (t,2H); 4.2 (q, 2H); 5.1 (s, 2H); 7.0 (d, 2H); 7.2-7.3 (m, 3H); 7.4 (m,1H); 8.0 (d, 2H).

Example 17 Synthesis of Ethyl4-(4-(2,6-difluorobenzyloxyl)phenyl)-4-oxobutyrate

Step A: Preparation of 4-(2,6-Difluorobenzyloxy)acetophenone

Using the method of Example 8, Step A, the title compound was obtained

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 4H);7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyrate

To a stirred solution of 4-(2,6-Difluorobenzyloxy)acetophenone (Step A,0.6 g, 22.8 mmol) in dry THF (60 ml) and DMPU (12 ml) was added asolution of lithium bis(trimethylsilyl)amide (1.0M, 30 ml) at −60° C.under argon. After stirring for 10 minutes at −60° C., Ethylbromoacetate (7.61 g, 45.6 mmol) was added rapidly. The reaction mixturewas stirred for an additional 10 minutes and then warmed to roomtemperature for 4 hours. The crude mixture was taken in EtOAc and washedwith water and brine. The aqueous layer was extracted one more time withEtOAc. The combined organic layers were dried over Na₂SO₄, filtered,concentrated and purified by flash chromatography on silica gel column(hex:ethyl acetate, 4:1) to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 1.3 (t, 3H); 2.8 (t, 3H); 3.2 (t, 2H); 4.1 (q,2H); 5.2 (s, 2H); 6.9-7.0 (m, 4H); 7.3-7.4 (m, 1H); 7.9 (d, 2H).

Example 18 Synthesis of 4-(4-(2-(2-Thienyl)ethoxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2-(2-Thienyl)ethoxy)acetophenone

Using the method of Example 2, Step A, using 2-(2-Thienyl)ethanol as thestarting material and purification by flash chromatography on silica gelcolumn (hex:ethylacetate, 3:1), the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 3.3 (t, 2H); 4.2 (t, 2H); 6.9-7.1(m, 4H); 7.2 (d, 1H); 7.9 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2-(2-thienyl)ethoxy)phenyl)-4-oxobutyrate

Using the method of Example 2, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.3 (t, 3H); 2.8 (t, 2H); 3.3 (m, 4H); 4.1 (q,2H); 4.2 (t, 2H); 6.9-7.1 (m, 4H); 7.2 (d, 1H); 7.9 (d, 2H).

Step C: Preparation of 4-(4-(2-(2-Thienyl)ethoxy)phenyl)-4-oxobutyricacid

Using the method of Example 2, Step C, the title compound was obtained

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.3 (m, 4H); 4.2 (t, 2H); 6.9-7.1(m, 4H); 7.2 (d, 1H); 7.9 (d, 2H).

Example 19 Synthesis of 4-(2,6-Difluorophenyl)-4-oxobutyric acid

Step A: Preparation of tert-Butyl 4-(2,6-difluorophenyl)-4-oxobutyrate

To a stirred solution of 2,6-Difluoroacetophenone (5 g, 32 mmol) in dryTHF (40 ml) and DMPU (8 ml) was added a solution of lithiumbis(trimethylsilyl)amide (1.0M, 45 ml) at −60° C. under argon. Afterstirring for 10 minutes at −60° C., tert-Butyl bromoacetate (6.99 g,35.8 mmol) was added rapidly. The reaction mixture was stirred for anadditional 10 minutes and then warmed to room temperature for 4 hours.The crude mixture was taken in EtOAc and washed with water and brine.The aqueous layer was extracted one more time with EtOAc. The combinedorganic layers were dried over Na₂SO₄, filtered, concentrated andpurified by flash chromatography on a silica gel column (hex:ethylacetate, 2:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.8 (t, 2H); 3.2 (t, 2H); 6.9-7.0(m, 2H); 7.4 (m, 1H).

Step B: Preparation of Compound AS

A solution of tert-Butyl 4-(2,6-difluorophenyl)-4-oxobutyrate (Step A,9.52 g, 35.2 mmol) in dichloromethane (30 ml) was treated withtrifluoroacetic acid (20 ml). The mixture was stirred at ambienttemperature for 3 hours and concentrated. The purification was done byflash chromatography on silica gel column (chloroform:methanol, 95:5spiked with acetic acid) to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 6.9-7.0 (m, 2H); 7.4(m, 1H).

Example 20 Synthesis of 4-(4-(2,5-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2,5-Dimethylbenzyloxy)acetophenone

Using the method of Example 8, Step A, using 2,5-Dimethylbenzyl chlorideas the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 3H); 2.5 (s, 3H); 5.1 (s, 2H); 6.9-7.2(m, 5H); 7.9 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2,5-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.3 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.0 (d, 2H); 7.2-7.3 (m, 3H); 7.9 (d,2H).

Step C: Preparation of 4-(4-(2,5-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

To a solution of Ethyl 4-(4-(2,5-dimethylbenzyloxy)phenyl)-4-oxobutyrate(Step B, 2.62 g, 7.7 mmol) in abs ethanol (30 ml) was added 1N NaOH (10ml) at room temperature. The reaction mixture was stirred for 3 hoursand then acidified with 1M HCl. The occurring white precipitate wasfiltered, washed with water and dried under vacuum to provide the titlecompound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 6H); 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s,2H); 7.0 (d, 2H); 7.2-7.3 (m, 3H); 8.0 (d, 2H).

Example 21 Synthesis of 4-(4-(2,5-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2,5-Difluorobenzyloxy)acetophenone

Using the method of Example 8, Step A, using 2,5-Difluorolbenzyl bromideas the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.1 (s, 2H); 6.9-7.0 (m, 3H); 7.2(m, 2H); 8.0 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2,5-difluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.8 (t, 2H); 3.2 (t, 2H); 4.4 (q,2H); 5.1 (s, 2H); 6.9-7.0 (m, 3H); 7.2 (m, 2H); 8.0 (d, 2H).

Step C: Preparation of 4-(4-(2,5-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

To a solution of Ethyl 4-(4-(2,5-Difluorobenzyloxy)phenyl)-4-oxobutyrate(Step B, 16.51 g, 47.4 mmol) in abs ethanol (100 ml) was added 1N NaOH(40 ml) at room temperature. The reaction mixture was stirred for 3hours, acidified with 1M HCl and concentrated in vacuo. The crudemixture was taken in chloroform and washed with water. The aqueous layerwas washed one more time with chloroform. The combined organic layerswere dried over Na₂SO₄, filtered and concentrated. The purification wasdone by flash chromatography on silica gel column (chloroform:methanol,95:5 spiked with acetic acid) to provide the title compound as a whitesolid.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s, 2H); 6.9-7.0(m, 3H); 7.2 (m, 2H); 8.0 (d, 2H).

Example 22 Synthesis of 4-(4-(2,4-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-(2,4-Difluorobenzyloxy)acetophenone

Using the method of Example 8, Step A, using 2,4-Difluorolbenzyl bromideas the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.1 (s, 2H); 6.9-7.0 (m, 2H); 7.1(d, 2H); 7.4 (m, 1H); 8.0 (d, 2H).

Step B: Preparation of Ethyl4-(4-(2,4-difluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.8 (t, 2H); 3.2 (t, 2H); 4.4 (q,2H); 5.1 (s, 2H); 6.9-7.0 (m, 2H); 7.1 (d, 2H); 7.4 (m, 1H); 8.0 (d,2H).

Step C: Preparation of 4-(4-(2,4-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 21, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s, 2H); 6.9-7.0(m, 2H); 7.1 (d, 2H); 7.4 (m, 1H); 8.0 (d, 2H).

Example 23 Synthesis of 4-(3-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 3-(2,6-Difluorobenzyloxy)acetophenone

Using the method of Example 8, Step A, using 3-Hydroxyacetophenone asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 2H); 7.2(m, 1H); 7.4 (m, 2H); 7.9 (d, 2H).

Step B: Preparation of Ethyl4-(3-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.8 (t, 2H); 3.2 (t, 2H); 4.4 (q,2H); 5.1 (s, 2H); 6.9-7.0 (m, 2H); 7.2 (m, 1H); 7.4 (m, 2H); 7.9 (d,2H).

Step C: Preparation of 4-(3-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 21, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.1 (s, 2H); 6.9-7.0(m, 2H); 7.2 (m, 1H); 7.4 (m, 2H); 7.9 (d, 2H).

Example 24 Synthesis of 4-(4-((Cyclopropyl)-methoxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 4-((Cyclopropyl)-methoxy)acetophenone

Using the method of Example 8, Step A, using Cyclopropylmethyl bromideas the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 0.4 (m, 2H); 0.6 (m, 2H); 1.2 (m, 1H); 2.5 (s,3H); 3.8 (d, 2H); 6.9 (d, 2H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-((cyclopropyl)-methoxy)phenyl)-4-oxobutyrate

Using the method of Example 8, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 0.4 (m, 2H); 0.6 (m, 2H); 1.2 (m, 1H); 1.4 (s,9H); 2.6 (t, 2H); 3.2 (t, 2H); 3.8 (d, 2H); 6.9 (d, 2H); 7.9 (d, 2H).

Step C: Preparation of 4-(4-((Cyclopropyl)-methoxy)phenyl)-4-oxobutyricacid

Using the method of Example 8, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 0.4 (m, 2H); 0.6 (m, 2H); 1.2 (m, 1H); 2.8 (t,2H); 3.2 (t, 2H); 3.8 (d, 2H); 6.9 (d, 2H); 7.9 (d, 2H).

Example 25 Synthesis of4-(4-(2-Trifluoromethylbenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(2-Trifluoromethylbenzyloxy)acetophenone

Using the method of Example 8, Step A, using 2-(Trifluoromethyl)benzylbromide as the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.3 (s, 2H); 6.9 (d, 2H); 7.4 (t,1H); 7.6 (t, 1H); 7.7 (d, 2H); 7.9 (d, 2H).

Step B: Preparation of tert-Butyl4-(4-(2-trifluoromethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 8, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 2.7 (t, 2H); 3.2 (t, 2H); 5.3 (s,2H); 6.9 (d, 2H); 7.4 (t, 1H); 7.6 (t, 1H); 7.7 (d, 2H); 7.9 (d, 2H).

Step C: Preparation of4-(4-(2-Trifluoromethylbenzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 8, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.2 (t, 2H); 5.3 (s, 2H); 6.9 (d,2H); 7.4 (t, 1H); 7.6 (t, 1H); 7.7 (t, 2H); 7.9 (d, 2H).

Example 26 Synthesis of3-[(4-(2,6-Difluorobenzyloxy)phenyl)-methylthio]propionic acid

Step A: Preparation of 4-Hydroxybenzyl bromide

Using the method of Example 3, Step A, the title compound was obtainedwhich was used without further purification.

Step B: Preparation of Ethyl 3-[(4-hydroxyphenyl)-methylthio]propionate

Using the method of Example 3, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.4-2.6 (m, 4H); 3.6 (s, 2H); 4.1(q, 2H); 6.7 (d, 2H); 7.2 (d, 2H).

Step C: Preparation of Ethyl 3-[(4-(2,6-difluorobenzyloxy)phenyl)methylthio]propionate

Using the method of Example 3, Step C, using 2,6-Difluorobenzyl bromideas the starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.4-2.6 (m, 4H); 3.6 (s, 2H); 4.2(q, 2H); 5.15 (s, 2H); 6.9 (d, 4H); 7.2-7.4 (m, 3H).

Step D: Preparation of3-[(4-(2,6-Difluorobenzyloxy)phenyl)-methylthio]propionic acid

Using the method of Example 3, Step D, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5-2.6 (m, 4H); 3.7 (s, 2H); 5.1 (s, 2H);6.9-7.0 (m, 4H); 7.2-7.4 (m, 3H).

Example 27 Synthesis of 4-(2-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 2-(2,6-Difluorobenzyloxy)acetophenone

Using the method of Example 8, Step A, using 2-Hydroxyacetophenone asthe starting material, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 3H); 7.1(d, 1H); 7.4 (m, 1H); 7.5 (t, 1H); 7.8 (d, 1H).

Step B: Preparation of Ethyl4-(2-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.6 (t, 2H); 3.2 (t, 2H); 4.1 (q,2H); 5.2 (s, 2H); 6.9-7.0 (m, 3H); 7.1 (d, 1H); 7.4 (m, 1H); 7.5 (t,1H); 7.8 (d, 1H).

Step C: Preparation of 4-(2-(2,6-Difluorobenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 21, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.6 (t, 2H); 3.2 (t, 2H); 5.2 (s, 2H); 6.9-7.0(m, 3H); 7.1 (d, 1H); 7.4 (m, 1H); 7.5 (t, 1H); 7.8 (d, 1H).

Example 28 Synthesis of Ethyl4-(4-(2,6-difluorobenzyloxy)phenyl)-3-oxobutyrate

Step A: Preparation of Ethyl 4-hydroxybenzylate

To a stirred solution of 4-Hydroxybenzyl alcohol (4 g, 26.28 mmol) indry DMF (15 ml), pyridine (1 ml) and N,N-Dicyclohexylcarbodiimide (6.50g, 31.5 mmol) was added abs EtOH (3.26 g, 78.84 mmol). The reactionmixture was stirred for 18 hours at room temperature and then filtered.The filtrate was concentrated under reduced pressure and purified byflash chromatography on silica gel column (hex:ethyl acetate, 2:1) toprovide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 3.5 (s, 2H); 4.1 (q, 2H); 6.7 (d,2H); 7.1 (d, 2H).

Step B: Preparation of Ethyl 4-(2,6-difluorobenzyloxy)benzylate

To a solution of NaH (60% dispersed in oil, 0.393 g, 9.8 mmol) in dryDMF (20 ml) was added Ethyl 4-hydroxybenzylate (Step A, 1.59 g, 8.8mmol). When the evolution of hydrogen ceased, 2,6-Difluorobenzyl bromide(1.64 g, 7.9 mmol) was added. The reaction mixture was stirred for 4hours at room temperature, quenched with sat. NH₄Cl, and concentrated invacuo. The residue was taken in EtOAc and washed twice with water andbrine. The organic layer was dried over Na₂SO₄, filtered, concentratedand purified by flash chromatography on silica gel column (hex:ethylacetate, 2:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 3.5 (s, 2H); 4.1 (q, 2H); 5.1 (s,2H); 6.9 (m, 4H); 7.2-7.4 (m, 3H).

Step C: Preparation of 4-(2,6-Difluorobenzyloxy)benzylic acid

To a stirred solution of Ethyl 4-(2,6-difluorobenzyloxy)benzylate (StepB, 2.14 g, 6.9 mmol) in abs EtOH (30 ml) was added 1N NaOH (10 ml) atroom temperature. The reaction mixture was stirred for 3 hours,acidified by 1M HCl and filtered. The white precipitate was washed withwater and dried under high vacuum to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 3.6 (s, 2H); 5.1 (s, 2H); 6.9 (m, 4H); 7.2-7.4(m, 3H).

Step D: preparation of 4-(2,6-Difluorobenzyloxy)benzylcarbonyl chloride

Thionyl chloride (10 ml) was added to 4-(2,6 Difluorobenzyloxy)benzylicacid (Step C, 1.61 g, 5.79 mmol). The reaction mixture was refluxed for3 hours and concentrated in vacuo to provide light yellow oil which wasused without further purification.

Step E: Ethyl 4-(4-(2,6-difluorobenzyloxy)phenyl)-3-oxobutyrate

To a solution of Meldrum's acid (0.846 g, 5.8 mmol) in dichloromethane(5 ml) was added pyridine (2 ml) over a period of 10 minutes at 0° C. Tothis solution was added 4-(2,6-Difluorobenzyloxy)benzylcarbonyl chloride(Step D, 1.71 g, 5.7 mmol) in dichloromethane (5 ml), which resulted inan orange solution. The dark orange solution was stirred for 1 hour at0° C., allowed to warm to room temperature and stirred for an additionalhour. The reaction mixture was diluted with dichloromethane and pouredonto 2M HCl and ice. The phases were separated and the aqueous phase wasextracted twice with dichloromethane. The combined organic layers werewashed twice with 2M HCl and brine, dried over Na₂SO₄, filtered andconcentrated to a solid. The solid was suspended in abs EtOH (15 ml) andrefluxed for 2.5 hours. The solvent was removed in vacuo to give a darkoil. The residue was purified by flash chromatography on silica gelcolumn (hex:ethyl acetate, 2:1) to provide the title compound as a whitesolid.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 3.4 (s, 2H); 3.7 (s, 2H); 4.2 (q,2H); 5.1 (s, 2H); 6.9 (m, 4H); 7.1 (d, 2H); 7.3 (m, 1H).

Example 29 Synthesis of3-(2-(4-(2,6-Difluorobenzyloxy)phenyl)-2-oxoethyl)thio-1H-1,2,4-triazole

Step A: Preparation of 4-(2,6-Difluorobenzyloxy)acetophenone

Using the method of Example 8, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 4H);7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of2-Bromo-1-(4-(2,6-difluorobenzyloxy)phenyl)-1-Ethanone

To a stirred solution of copper (2) bromide (3.70 g, 16.6 mmol) in ethylacetate (20 ml) was added a solution of4-(2,6-Difluorobenzyloxy)acetophenone (Step A, 2.74 g, 10.4 mmol) inchloroform (20 ml) at room temperature. The reaction mixture wasrefluxed for 16 hours and then water was added. The crude mixture wasextracted twice with EtOAc. The organic layers were combined and washedwith water, brine, dried over Na₂SO₄, filtered, concentrated andpurified by flash chromatography on silica gel column (hex:ethylacetate, 4:1) to provide the title compound as a white flaky solid.

¹H NMR (270 MHz, CDCl₃): 4.4 (s, 2H); 5.2 (s, 2H); 6.9-7.1 (m, 4H), 7.3(m, 1H); 8.0 (d, 2H).

Step C: Preparation of3-(2-(4-(2,6-Difluorobenzyloxy)phenyl)-2-oxoethyl)thio-1H-1,2,4-triazole

To a solution of 1H-1,2,4-Triazole-3-thiol (0.250 g, 2.4 mmol) andtriethylamine (2.50 g, 2.4 mmol) in dry dichloromethane (20 ml) wasadded 2-Bromo-1-(4-(2,6-difluorobenzyloxy)phenyl)-1-Ethanone (Step B,0.851 g, 2.4 mmol) in dry dichloromethane (5 ml) at room temperature.The reaction mixture was stirred for 50 minutes and then concentrated invacuo. The crude residue was taken in EtOAc and washed with 0.1M HCl,and brine. The organic layer was dried over Na₂SO₄, filtered,concentrated and purified by flash chromatography on silica gel column(chloroform:methanol, 9:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 4.5 (s, 2H); 5.1 (s, 2H); 6.8-7.0 (m, 4H), 7.2(m, 1H); 7.9 (d, 2H); 8.0 (s, 1H).

Example 30 Synthesis of5-((4-(2,6-Difluorobenzyloxy)phenyl)-methyl)-1H-tetrazole

Step A: Preparation of (4-(2,6-Difluorobenzyloxy)phenyl)-acetonitrile

To a solution of 4-Hydroxybenzyl cyanide (5 g, 37.5 mmol) and K₂CO₃(6.74 g, 48.8 mmol) in dry DMF (20 ml) was added 2,6-Difluorobenzylbromide (7.77 g, 37.5 mmol). The reaction mixture was stirred for 4hours at room temperature and concentrated in vacuo. The crude residuewas taken in EtOAc and washed with water and brine. The aqueous layerwas washed one more time with EtOAc. The combined organic layers weredried over Na₂SO₄, filtered, and concentrated to provide the titlecompound as a white solid.

¹H NMR (270 MHz, CDCl₃): 3.65 (s, 2H); 5.1 (s, 2H); 6.9-7.0 (m, 4H);7.2-7.4 (m, 3H);

Step B: Preparation of5-((4-(2,6-Difluorobenzyloxy)phenyl)-methyl)-1H-tetrazole

A mixture of (4-(2,6-Difluorobenzyloxy)phenyl)-acetonitrile (Step A, 5g, 19.3 mmol), NaN₃ (1.3 g, 20 mmol), and NH₄Cl (1.06 g, 20 mmol) in dryDMF (60 ml) was heated at 90° C. for 16 hours. The solvent was removedin vacuo and the oily residue was partitioned between EtOAc and water(acidified to pH 1 with conc. HCl). The organic layer was washed withwater, dried over Na₂SO₄, filtered and concentrated to a brownsemisolid. The purification was done by flash chromatography on silicagel column (chloroform:methanol, 9:1) to provide the title compound as alight creamy solid.

¹H NMR (270 MHz, CDCl₃): 4.0 (s, 2H); 5.1 (s, 2H); 6.7-6.9 (m, 4H); 7.0(d, 2H); 7.2 (m, 1H).

Example 31 Synthesis of (2RS)2-(N-Boc)-3-[2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl]thiopropionic acid

Step A: Preparation of 4-(2,6-Difluorobenzyloxy)acetophenone

To a solution of 4-Hydroxyacetophenone (3.28 g, 24 mmol) and K₂CO₃ (4.33g, 31.3 mmol) in dry DMF (15 ml) was added 2,6-Difluorobenzyl bromide (5g, 24.1 mmol). The reaction mixture was stirred at room temperature for5 hours, quenched with water and concentrated in vacuo. The cruderesidue was taken in EtOAc and washed with water and brine. The aqueouslayer was extracted twice with EtOAc. The combined organic layers weredried over Na₂SO₄, filtered, concentrated and purified by flashchromatography on silica gel column (hex:ethyl acetate, 2:1) to providethe title compound.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 4H);7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of 2-Bromo-1-(4-(2,6-difluorobenzyloxy)phenyl)-1-Ethanone

Using the method of Example 29, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 4.4 (s, 2H); 5.2 (s, 2H); 6.9-7.1 (m, 4H), 7.3(m, 1H); 8.0 (d, 2H).

Step C: Preparation of Ethyl(2RS)2-(N-Boc)-3-[2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl]thiopropionate

To a stirred solution of2-Bromo-1-(4-(2,6-difluorobenzyloxy)phenyl)-1-Ethanone (Step B, 2.07 g,8.3 mmol) in dry dichloromethane (20 ml) and triethylamine (8.39 g, 83mmol) was added Boc-Cys-OEt (2.94 g, 8.6 mmol). The reaction mixture wasstirred at room temperature for an hour and concentrated in vacuo. Thecrude residue was taken in EtOAc, washed with 0.1M HCl, and brine. Theorganic layer was dried over Na₂SO₄, filtered, concentrated and purifiedby flash chromatography on silica gel column (chloroform:methanol,97.5:2.5) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 1.4 (s, 9H); 3.0 (m, 2H); 3.8 (s,2H); 4.2 (q, 2H); 4.5 (br, 1H); 5.2 (s, 2H); 5.4 (d, 1H); 6.9-7.1 (m,4H); 7.3 (m, 1H); 7.9 (d, 2H).

Step D: Preparation of(2RS)2-(N-Boc)-3-[2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl]thiopropionicacid

To a solution of Ethyl(2RS)2-(N-Boc)-3-[2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl]thiopropionate(Step C, 0.761 g, 1.5 mmol) in abs EtOH (10 ml) was added 1N NaOH (3ml). The reaction mixture was stirred at room temperature for 4 hours,acidified with 1M HCl and concentrated in vacuo. The crude residue wastaken in chloroform, and washed with water and brine. The organic layerwas dried over Na₂SO₄, filtered, concentrated and purified by flashchromatography on silica gel column (chloroform:methanol, 92.7:7.5) toprovide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.4 (s, 9H); 3.0 (t, 2H); 4.0 (q, 2H); 4.5 (br,1H); 5.2 (s, 2H); 5.4 (d, 1H); 6.9-7.1 (m, 4H); 7.3 (m, 1H); 7.9 (d,2H).

Example 32 Synthesis of Ethyl2-Hydroxy-4-oxo-4-(4-(2,6-difluorobenzyloxy)phenyl)but-2-enoate

Step A: Preparation of 4-(2,6-Difluorobenzyloxy)acetophenone

Using the method of example 31, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 4H);7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of Ethyl2-Hydroxy-4-oxo-4-(4-(2,6-difluorobenzyloxy)phenyl)but-2-enoate

A mixture of 4-(2,6-Difluorobenzyloxy)acetophenone (Step A, 5.64 g, 21.5mmol) and diethyl oxalate (3.14 g, 21.5 mmol) was added to an ice-cooledsolution of NaOEt (0.490 g, 22.4 mmol of metallic Na) in abs EtOH (25ml). After being allowed to stand overnight at room temperature, themixture was diluted with water (50 ml), acidified with 10% HCl, andextracted thrice with EtOAc. The combined organic layers were washedwith brine, dried over Na₂SO₄, filtered, concentrated and purified byflash chromatography on silica gel column (hex:ethyl acetate, 4:1) toprovide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.4 (t, 3H); 4.4 (q, 2H); 5.2 (s, 2H); 6.9-7.1(m, 5H); 7.3-7.4 (m, 1H); 8.0 (d, 2H).

Example 33 Synthesis of (2RS)2-(N-Acetyl)-4-(4-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 4-(2,6-Difluorobenzyloxy)acetophenone

Using the method of example 31, Step A, the title compound was obtained.¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.2 (s, 2H); 6.9-7.0 (m, 4H);7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step B: Preparation of 2-Bromo-1-(4-(2,6-difluorobenzyloxy)phenyl)-1-Ethanone

Using the method of Example 29, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 4.4 (s, 2H); 5.2 (s, 2H); 6.9-7.1 (m, 4H), 7.3(m, 1H); 8.0 (d, 2H).

Step C: Preparation of Diethyl(N-Acetyl)(2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl)propanedioate

To a solution of Diethyl acetamidomalonate (0.949 g, 4.3 mmol) and NaOEt(0.301 g, 4.4 mmol) in abs EtOH (25 ml) was added2-Bromo-1-(4-(2,6-difluorobenzyloxy) phenyl)-1-Ethanone (Step B, 1.42 g,4.1 mmol). The reaction mixture was stirred at room temperature for 2hours and concentrated in vacuo. The crude residue was partitionedbetween EtOAc and 0.01 N NaOH. The organic layer was washed with waterand 0.001 M HCl, dried over Na₂SO₄, filtered and concentrated. Thepurification was done by flash chromatography on silica gel column(hex:ethyl acetate, 2:1) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 6H); 2.0 (s, 3H); 4.2-4.3 (m, 6H); 5.2(s, 2H); 6.9-7.1 (m, 4H), 7.3-7.4 (m, 1H); 7.9 (d, 2H).

Step D: Preparation of(2RS)2-(N-Acetyl)-4-(4-(2,6-difluorobenzyloxy)phenyl)-4-oxobutyric acid

To a solution of Diethyl(N-Acetyl)(2-(4-(2,6-difluorobenzyloxy)phenyl)-2-oxoethyl)propanedioate(Step C, 1.28 g, 2.6 mmol) in water (20 ml) was added NaOH (0.529 g,13.2 mmol). The reaction mixture was refluxed for 16 hours, then glacialacetic acid (18 ml) was added and refluxing continued for an additional3 hours. The mixture was concentrated in vacuo and purified by flashchromatography on silica gel column (chloroform:methanol, 9:1) toprovide the title compound.

¹H NMR (270 MHz, CDCl₃:CD₃OD): 2.0 (s, 3H); 3.5 (m, 2H); 4.8 (t, 1H);5.1 (s, 2H); 6.9-7.1 (m, 4H), 7.3-7.4 (m, 1H); 7.9 (d, 2H).

Example 34 Synthesis of 4-(3-((Cyclopropyl)-methoxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 3-((Cyclopropyl)-methoxy)acetophenone

Using the method of Example 31, Step A, using Cyclopropylmethyl bromideand 3-Hydroxyacetophenone as the starting materials, the title compoundwas obtained.

¹H NMR (270 MHz, CDCl₃): 0.4 (m, 2H); 0.6 (m, 2H); 1.2 (m, 1H); 2.5 (s,3H); 3.8 (d, 2H); 7.1 (m, 1H); 7.4 (m, 1H); 7.5-7.6 (m, 2H).

Step B: Preparation of tert-Butyl4-(3-((cyclopropyl)-methoxy)phenyl)-4-oxobutyrate

Using the method of Example 8, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 0.4 (m, 2H); 0.6 (m, 2H); 1.2 (m, 1H); 1.4 (s,9H); 2.6 (t, 2H); 3.2 (t, 2H); 3.8 (d, 2H); 7.1 (m, 1H); 7.4 (m, 1H);7.5-7.6 (m, 2H).

Step C: Preparation of 4-(3-((Cyclopropyl)-methoxy)phenyl)-4-oxobutyricacid

Using the method of Example 8, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 0.4 (m, 2H); 0.6 (m, 2H); 1.2 (m, 1H); 2.8 (t,2H); 3.2 (t, 2H); 3.8 (d, 2H); 7.1 (m, 1H); 7.4 (m, 1H); 7.5-7.6 (m,2H).

Example 35 Synthesis of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

To a solution of 2,6-dimethylbenzoic acid (10 g, 66.5 mmol) andpotassium carbonate (9.18 g, 66.5 mmol) in dimethylformamide (67 ml),was added methyl iodide (8.28 ml, 133.16 mmol) in an ice bath, and themixture was stirred for 16 hours. To the reaction mixture was addedtoluene and water, and the organic layer was washed with 3% K₂CO₃, 1NHCl, and brine. The organic layer was dried over Na₂SO₄, filtered andconcentrated.

The oily residue was redissolved in dry THF (135 ml), added to LiAlH₄(3.79 g, 99.8 mmol), and stirred for 4 hours in an ice bath. To thereaction mixture was added 1N HCl slowly followed by ethyl acetate, andthe organic layer was washed with brine, dried over Na₂SO₄, filtered andconcentrated. The oily residue was used without further purification.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

To a stirred solution of 3′-Hydroxyacetophenone (8.07 g, 59.24 mmol) andTriphenylphosphine (16.93 g, 64.5 mmol) in dry THF (180 ml) was addeddropwise 2,6-Dimethylbenzyl alcohol (8.05 g, 59.24 mmol) and diethylazodicarboxylate (11.24 g, 64.57 mmol) in dry THF (45 ml) and dry DMF(18 ml) at ambient temperature. After stirring for 1.5 hours at ambienttemperature, the reaction mixture was diluted with ether and washedtwice with water, 1N NaOH and brine, dried over Na₂SO₄, filtered andconcentrated. The purification was done by flash chromatography onsilica gel column (hex:ethyl acetate, 2:1) to provide the titlecompound.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

To a solution of Ethyl 4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate(Step C, 12.31 g, 36.2 mmol) in abs ethanol (160 ml) was added 1N NaOH(50 ml) at room temperature. The reaction mixture was stirred for 3hours and then acidified with 1M HCl. The occuring white precipitate wasfiltered, washed with water and dried under vacuum to provide the titlecompound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Example 36 Synthesis of4-(3-(2-Fluoro-6-methylbenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 2-Fluoro-6-methylbenzoic Acid

Synthesized as described in Example 89(d) of International PatentPublication No. WO 97/34893, page 43.

Step B: Preparation of 2-Fluoro-6-methylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 4.7 (s, 2H); 6.85 (t, 1H); 6.95(d, 1H); 7.15 (m, 1H).

Step C: Preparation of 3-(2-Fluoro-6-methylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (m,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step D: Preparation of Ethyl4-(3-(2-Fluoro-6-methylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 3H); 2.8 (t, 2H); 3.3 (t,2H); 4.4 (q, 2H); 5.2 (s, 2H); 6.9-7.1 (m, 2H); 7.2 (m, 2H); 7.4 (t,1H); 7.6 (m, 2H).

Step E: Preparation of4-(3-(2-Fluoro-6-methylbenzyloxy)phenyl)-4-oxobutyric acid

To a solution of Ethyl4-(3-(2-Fluoro-6-methylbenzyloxy)phenyl)-4-oxobutyrate (Step D, 8.56 g,24.9 mmol) in abs ethanol (100 ml) was added 1N NaOH (40 ml) at roomtemperature. The reaction mixture was stirred for 3 hours, acidifiedwith 1M HCl and concentrated. The residue was taken in chloroform andwashed with 0.1 M HCl, brine, dried over Na₂SO₄, filtered andconcentrated. The purification was done by flash chromatography onsilica gel column (chloroform:methanol 95:5 spiked with acetic acid) toprovide the title compound as white solid.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 3H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 6.9-7.1 (m, 2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Example 37 Synthesis of Ethyl4-(3-(2,6-dimethylbenzyloxyl)phenyl)-4-oxobutyrate

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Example 38 Synthesis of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid Sodium salt

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step E: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid Sodium salt

The 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid (Step D, 5.5g, 17.6 mmol) was dissolved in abs ethanol (20 ml) by warming gentlyfollowed by addition of NaOH (0.705 g) at 0° C. temperature. Thereaction mixture was stirred for one hour, concentrated in vaccuo andlyophilized to give as a white solid.

¹H NMR (270 MHz, D₂O): 2.0 (s, 6H); 2.5 (t, 2H); 3.0 (t, 2H); 4.8 (s,2H); 6.8 (d, 2H); 6.9 (m, 2H); 7.2 (t, 1H); 7.5 (d, 2H).

Example 39 Synthesis of 4-(4-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 4-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.0-7.2(m, 5H); 8.0 (d, 2H).

Step C: Preparation of Ethyl4-(4-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.0-7.2 (m, 5H); 8.0 (d, 2H).

Step D: Preparation of 4-(4-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.0-7.2 (m, 5H); 8.0 (d, 2H).

Example 40 Synthesis of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid Potassium salt

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.3 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (d,2H); 7.2 (m, 2H); 7.45 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.45 (t, 1H);7.6 (m, 2H).

Step D: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.5 (t, 2H); 3.2 (t, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.45 (t, 1H); 7.6 (m, 2H).

Step E: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid Potassium salt

The 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid (Step D, 6.0g, 19.4 mmol) was dissolved in abs ethanol (20 ml) by warming gentlyfollowed by addition of KOH (1.21 g) at 0° C. temperature. The reactionmixture was stirred for one hour, concentrated in vaccuo and lyophilizedto give the title compound as white solid.

¹H NMR (270 MHz, D₂O): 2.3 (s, 6H); 2.5 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.45 (t, 1H); 7.6 (m, 2H).

Example 41 Synthesis of4-(3-(2,6-Dimethoxybenzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 2,6-Dimethoxylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 3.9 (s, 6H); 4.8 (s, 2H); 6.5 (d, 2H); 7.25 (m,1H).

Step B: Preparation of 3-(2,6-Dimethoxybenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.6 (s, 3H); 3.9 (s, 6H); 5.2 (s, 2H); 6.6 (d,2H); 7.3 (m, 3H); 7.5 (d, 1H); 7.7 (d, 1H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethoxybenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.8 (t, 2H); 3.3 (t, 2H); 3.8 (s,6H); 4.1 (q, 2H); 5.2 (s, 2H); 6.5 (d, 2H); 7.3-7.4 (m, 3H); 7.6 (d,1H); 7.7 (d, 1H).

Step D: Preparation of 4-(3-(2,6-dimethoxybenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.8 (t, 2H); 3.3 (t, 2H); 3.8 (s, 6H); 5.2 (s,2H); 6.5 (d, 2H); 7.3-7.4 (m, 3H); 7.6 (d, 1H); 7.7 (d, 1H).

Example 42 Synthesis of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxo-2,2-dimethylbutyric acid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxo-2,2-dimethylbutyrate

To a stirred solution of 3-(2,6-Dimethylbenzyloxy)acetophenone (Step B,4.11 g, 16.1 mmol) in dry THF (60 ml) and DMPU (12 ml) was added asolution of lithium bis(trimethylsilyl)amide (1.0M, 17.74 ml) at −60° C.under argon. After stirring for 10 minutes at −60° C., Ethyl2-bromoisobutyrate (4.73 g, 24.2 mmol) was added rapidly. The reactionmixture was stirred for an additional 10 minutes and then warmed to roomtemperature for 4 hours. The crude mixture was taken in EtOAc and washedwith water. The aqueous layer was extracted one more time with EtOAc.The combined organic layers were washed with brine, dried over Na₂SO₄,filtered, concentrated and purified by flash chromatography on silicagel column (hex:ethyl acetate, 4:1) to provide the title compound aswhite solid.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 1.3 (s, 6H); 2.3 (s, 6H); 3.3 (s,2H); 4.1 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxo-2,2-dimethylbutyric acid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.3 (s, 6H); 2.3 (s, 6H); 3.3 (s, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Example 43 Synthesis of4-(3-(4-(Trifluoromethyl)benzyloxy)phenyl)-4-oxobutyric acid

Step A: Preparation of 3-(4-(Trifluoromethyl)benzyloxy)acetophenone

Using the method of Example 31, Step A, using 4-(Trifluoromethyl)benzylbromide and 3-Hydroxyacetophenone as the starting materials, the titlecompound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 3H); 5.1 (s, 2H); 7.1 (d, 2H); 7.4-7.6(m, 6H).

Step B: Preparation of Ethyl4-(3-(4-(trifluoromethyl)benzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.3 (t, 3H); 2.7 (t, 2H); 3.3 (t, 2H); 4.1 (q,2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.4-7.6 (m, 6H).

Step C: Preparation of4-(3-(4-(Trifluoromethyl)benzyloxy)phenyl)-4-oxobutyric acid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.7 (t, 2H); 3.3 (t, 2H); 5.1 (s, 2H); 7.1 (d,2H); 7.4-7.6 (m, 6H).

Example 44 Synthesis of 4-(3-((Cyclobutyl)-methoxy)phenyl)-4-oxobutyricacid

Step A: Preparation of 3-((Cyclobutyl)-methoxy)acetophenone

Using the method of Example 31, Step A, using Cyclobutylmethyl bromideand 3-Hydroxyacetophenone as the starting materials, the title compoundwas obtained.

¹H NMR (270 MHz, CDCl₃): 1.9 (m, 4H); 2.1 (m, 2H); 2.5 (s, 3H); 2.7 (m,1H); 4.0 (d, 2H); 7.1 (dd, 1H); 7.4 (t, 1H); 7.5-7.6 (m, 2H).

Step B: Preparation of Ethyl4-(3-((cyclobutyl)-methoxy)phenyl)-4-oxobutyrate

Using the method of Example 35, Step C, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 1.9 (m, 4H); 2.1 (m, 2H); 2.7 (m,1H); 2.8 (t, 2H); 3.3 (t, 2H); 4.0 (d, 2H); 4.1 (q, 2H); 7.1 (dd, 1H);7.4 (t, 1H); 7.5-7.6 (m, 2H).

Step C: Preparation of 4-(3-((Cyclobutyl)-methoxy)phenyl)-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.9 (m, 4H); 2.1 (m, 2H); 2.7 (m, 1H); 2.8 (t,2H); 3.3 (t, 2H); 4.0 (d, 2H); 7.1 (dd, 1H); 7.4 (t, 1H); 7.5-7.6 (m,2H).

Example 45 Synthesis of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)butyric acid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H): 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H): 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step E: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)butyric acid

A solution of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid(Step D, 3 g, 9.6 mmol), hydrazine (1.41 ml, 28.8 mmol) and potassiumhydroxide (1.61 g, 28.8 mmol) in ethylene glycol (12 ml) was refluxedfor 4 h, water (18 ml) and 6 N HCl (10 ml) were added to the reactionmixture. The crude reaction mixture was concentrated, and the residuewas dissolved in EtOAc, washed with water and brine, dried over Na₂SO₄,filtered, and concentrated. The purification was done by flashchromatography on silica gel column (chloroform:methanol 95:5 spikedwith acetic acid) to provide the title compound as a white solid.

¹H NMR (270 MHz, CDCl₃): 2.4 (m, 8H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Example 46 Synthesis of4-[[4-(2,6-Dimethylbenzyloxy)-3-methoxy]phenyl]-4-oxobutyric acid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 4-(2,6-Dimethylbenzyloxy)-3-methoxyacetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 3.9 (s, 3H); 5.1 (s,2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-[[4-(2,6-dimethylbenzyloxy)-3-methoxy]phenyl]-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t,2H); 3.9 (s, 3H); 4.4 (q, 2H); 5.1 (s, 2H); 7.0-7.2 (m, 4H); 7.6 (m,2H).

Step D: Preparation of4-[[4-(2,6-Dimethylbenzyloxy)-3-methoxy]phenyl]-4-oxobutyric acid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t, 2H); 3.9 (s,3H); 5.1 (s, 2H); 7.0-7.2 (m, 4H); 7.6 (m, 2H).

Example 47 Synthesis of4-{3-[((4-Trifluoromethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyricacid

Step A: Preparation of Methyl 2-methoxy-5-acetylbenzoate

To a stirred solution of Methyl 2-hydroxy-5-acetylbenzoate (12 g, 61.7mmol) in DMF (200 ml) was added cesium carbonate (24.15 g, 74.1 mmol)and MeI (9.64 g, 68 mmol).

The reaction mixture was stirred for 16 hours at 0° C. and then dilutedwith ethyl acetate, washed with aq Na₂S₂O₅, brine, dried over Na₂SO₄,filtered and concentrated. The purification was done by flashchromatography on silica gel column (ethyl acetate:hexane 1:2) toprovide the title compound as an off white solid.

¹H NMR (270 MHz, DMSO): 2.6 (s, 3H); 3.8 (s, 3H); 3.9 (s, 3H); 7.3 (d,1H); 8.1 (dd, 1H); 8.2 (s, 1H).

Step B: Preparation of 2-Methoxy-5-acetylbenzoic acid

Methyl 2-methoxy-5-acetylbenzoate (Step A, 3 g, 14.4 mmol) was dissolvedin acetic acid (80 ml) and then treated with c HCl (28 ml). The reactionmixture was refluxed for 4 hours, concentrated under reduced pressureand lyophilized to provide the title compound as cream color solid,which was used without further purification.

¹H NMR (270 MHz, DMSO): 2.6 (s, 3H); 3.9 (s, 3H); 7.3 (d, 1H); 8.1 (dd,1H); 8.2 (s, 1H).

Step C: Preparation of5-Acetyl-2-methoxy-N-[[4-(trifluoromethyl)phenyl]methyl]benzamide

To a stirred solution of 2-Methoxy-5-acetylbenzoic acid (Step B, 2.5 g,12.8 mmol), HOBt.H₂0 (2.08 g, 15.4 mmol), and EDC (3.70 g, 19.3 mmol) inCH₂Cl₂ (20 ml) and DMF (5 ml) was added 4-(Trifluoromethyl)benzylamine(2.48 g, 14.1 mmol), and the mixture was stirred for 16 hours at roomtemperature. The reaction mixture was concentrated under reducedpressure and then redissolved in ethyl acetate. The organic layer waswashed with 3% K₂CO₃, 1 N HCl, and brine, dried over Na₂SO₄, filteredand concentrated. The purification was done by flash chromatography onsilica gel column (chloroform:methanol 95:5) to provide the titlecompound as white solid.

¹H NMR (270 MHz, CDCl₃): 2.6 (s, 3H); 4.0 (s, 3H); 4.8 (d, 2H); 7.0 (d,1H); 7.5 (d, 2H); 7.6 (d, 2H); 8.1 (dd, 1H); 8.8 (s, 1H).

Step D: Preparation of Ethyl4-{3-[((4-Trifluoromethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.6 (t, 2H); 3.3 (t, 2H); 4.0 (s,3H); 4.4 (q, 2H); 4.8 (s, 2H); 7.0 (d, 1H); 7.4 (d, 2H); 7.6 (d, 2H);8.1 (dd, 1H); 8.8 (s, 1H).

Step E: Preparation of4-[5-[[N-(4-Trifluoromethylbenzyl)aminocarbonyl]-2-methoxy]phenyl]-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃: CD₃OD): 2.6 (t, 2H); 3.3 (t, 2H); 4.0 (s, 3H);4.7 (s, 2H); 7.0 (d, 1H); 7.4 (d, 2H); 7.6 (d, 2H); 8.1 (dd, 1H); 8.8(s, 1H).

Example 48 Synthesis of4-{3-[((2,6-Dimethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyricacid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of N-(2,6-Dimethylbenzyl)phthalimide

To a stirred solution of 2,6-Dimethylbenzyl Alcohol (Step A, 6.59 g,48.4 mmol) in DMSO (20 ml) was added chlorotrimethylsilane (15.75 ml,145 mmol) at room temperature, and the mixture was stirred for one hr.To this reaction mixture were added ethyl acetate and water, the organiclayer was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give an oil. The oily residue was redissolved in DMF(100 ml) and potassium phthalimide (10.76 g, 58.1 mmol) was added. Thereaction mixture was stirred for 16 hours at room temperature, ethylacetate was added, and the organic layer was washed with 3% Na₂CO₃, 1 NHCl, dried over Na₂SO₄, filtered and concentrated to give white solid.The purification was done by flash chromatography on silica gel column(chloroform:methanol 95:5) to provide the title compound as white solid.

¹H NMR (270 MHz, DMSO): 2.3 (s, 6H); 4.8 (s, 2H); 7.0 (m, 3H); 7.8 (s,4H).

Step C: Preparation of 2,6-Dimethylbenzyl Amine

To a stirred solution of N-(2,6-Dimethylbenzyl)phthalimide (Step B, 7.77g, 29.3 mmol) in ethanol (80 ml) was added hydrazine monohydrate (2.16ml, 44.52 mmol) and the reaction mixture was refluxed for 3.5 hours. Tothis reaction mixture was added c HCl to bring pH to 1 and refluxingcontinued for another 3.5 hours, water was added and reaction mixturewas filtered, the filtrate was concentrated and pH was adjusted to 10with 2 N NaOH. The residue was taken in methylene chloride and washedwith brine, dried over Na₂SO₄, filtered and concentrated to give an oilwhich was used without further purification.

¹H NMR (270 MHz, DMSO): 2.3 (s, 6H); 3.8 (s, 2H); 7.0 (m, 3H).

Step D: Preparation of5-Acetyl-2-methoxy-N-[[2,6-dimethyl)phenyl]methyl]benzamide

To a stirred solution of 2-Methoxy-5-acetylbenzoic acid (Example 47,Step B, 2.5 g, 12.8 mmol), HOBt (2.08 g, 15.4 mmol), and EDC (3.70 g,19.3 mmol) in CH₂Cl₂ (20 ml) and DMF (5 ml) was added 2,6-Dimethylbenzylamine (Step C, 1.72 g, 12.8 mmol), and the mixture was stirred for 16hours at room temperature. The reaction mixture was concentrated underreduced pressure and then redissolved in ethyl acetate. The organiclayer was washed with 3% K₂CO₃, 1 N HCl, and brine, dried over Na₂SO₄,filtered and concentrated. The purification was done by flashchromatography on silica gel column (chloroform:methanol 95:5) toprovide the title compound as white solid.

¹H NMR (270 MHz, CDCl₃): 2.5 (s, 6H); 2.6 (s, 3H); 3.9 (s, 3H); 4.7 (s,2H); 7.0 (d, 1H); 7.2 (m, 3H); 7.6 (br, 1H); 8.1 (dd, 1H); 8.8 (s, 1H).

Step E: Preparation of Ethyl4-{3-[((2,6-Dimethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (t, 3H); 2.4 (s, 6H); 2.7 (t, 2H); 3.3 (t,2H); 3.9 (s, 3H); 4.4 (q, 2H); 4.7 (s, 2H); 7.0 (m, 3H); 7.2 (m, 1H);8.1 (dd, 1H); 8.7 (s, 1H).

Step F: Preparation of4-{3-[((2,6-Dimethylbenzylamino)-carbonyl)-4-methoxy]phenyl}-4-oxobutyricacid

Using the method of Example 36, Step E, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃: CD₃OD): 2.4 (s, 6H); 2.7 (t, 2H); 3.3 (t, 2H);3.9 (s, 3H); 4.7 (s, 2H); 7.0 (m, 3H); 7.2 (m, 1H); 8.1 (dd, 1H); 8.7(s, 1H).

Example 49 Synthesis of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutanecarbohydroxamic acid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutanecarbohydroxamic acid

To a hydroxylamine hydrochloride solution in dry ethanol, add a solutionof potassium hydroxide in dry ethanol at 35° C. Cool the mix and addEthyl 4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate (Step C), andpowdered potassium hydroxide.

After few hours, reaction mixture can be diluted with water andneutralize with hydrochloric acid, filter and recrystallize to givetitle compound.

Example 50 Synthesis of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyramide

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyricacid

Using the method of Example 35, Step D, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.8 (t, 2H); 3.3 (t, 2H); 5.1 (s,2H); 7.1 (d, 2H); 7.2-7.3 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step E: Preparation of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyramide

To a solution of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxobutyric acid(Step D) in DMF, add triethylamine and BOP, after couple of hours ofstirring, reaction mixture can be added to liquid ammonia at −40° C. andthe resulting mixture can be warmed for 16 hours to give title compound.

Example 51 Synthesis of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxo-2-butenoic acid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxo-3-bromo-butyrate

To a ice cooled solution of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate (Step C, 3 g, 9 mmol)in dry ether (70 ml) was added dropwise bromine (0.7971 g, 9.9 mmol)diluted in ether (30 ml). After 4 hours of stirring, reaction mixturewas concentrated and purified by flash chromatography on silica gelcolumn (EtOAc:Hex, 1:4) to provide the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 3.1 (m, 1H); 3.5 (m,1H); 4.2 (q, 2H); 5.1 (s, 2H); 5.5 (m, 1H); 7.1 (d, 2H); 7.2 (m, 2H);7.4 (t, 1H); 7.6 (m, 2H).

Step E: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxo-2-butenoate

Triethylamine (5.95 g, 58.9 mmol) was added to a solution of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxo-3-bromo-butyrate (Step D, 2.47g, 5.8 mmol) in carbon tetrachloride (50 ml). After stirring for 4 hoursat room temperature, the reaction mixture was filtered through a pad ofsilica gel few times, and concentrated to give the title compound.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 4.2 (q, 2H); 5.1 (s,2H); 6.9 (dd, 1H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H);7.9 (dd, 1H).

Step F: Preparation of4-(3-(2,6-Dimethylbenzyloxy)phenyl)-4-oxo-2-butenoic acid

To a solution of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxo-2-butenoate (Step E) in absethanol at low temp, add aqueous sodium hydroxide, after an hour,concentrate and purify by flash chromatography on silica gel column(chloroform:methanol 95:5 spiked with acetic acid).

Example 52 Synthesis of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-3-butenoicacid

Step A: Preparation of 2,6-Dimethylbenzyl Alcohol

Using the method of Example 35, Step A, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 4.7 (s, 2H); 7.0-7.15 (m, 3H).

Step B: Preparation of 3-(2,6-Dimethylbenzyloxy)acetophenone

Using the method of Example 35, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 2.4 (s, 6H); 2.6 (s, 3H); 5.1 (s, 2H); 7.1 (dd,2H); 7.2 (m, 2H); 7.4 (t, 1H); 7.6 (m, 2H).

Step C: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate

Using the method of Example 17, Step B, the title compound was obtained.

¹H NMR (270 MHz, CDCl₃): 1.2 (s, 3H); 2.4 (s, 6H); 2.8 (t, 2H); 3.2 (t,2H); 4.4 (q, 2H); 5.1 (s, 2H); 7.1 (d, 2H); 7.2 (m, 2H); 7.4 (t, 1H);7.6 (m, 2H).

Step D: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-hydroxy-butyrate

To a solution of Ethyl 4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyrate(Step C) in tetrahydrofuran, add sodium borohydride dissolved in water.After 3-4 hours of stirring at room temperature, quench with an acid.The organic layer can be taken in dichloromethane, wash with water,aqueous sodium bicarbonate, and brine, dry over Na₂SO₄, filter andconcentrate. If needed, the compound can be purified by flashchromatography on silica gel column (EtOAc:Hex).

Step E: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-bromo-butyrate

To a solution of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-hydroxy-butyrate (Step D) indioxane, add phosphorous tribromide in dioxane dropwise. After stirringat room temperature for 16 hours, quench with water and chloroform.After few minutes, the reaction mixture can be neutralized by mildaqueous base, dry organic layer over Na₂SO₄, filter, concentrate andpurify by flash chromatography on silica gel column (EtOAc:hex).

Step F: Preparation of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-3-butenoate

Add Triethylamine to a solution of Ethyl4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-bromo-butyrate (Step E) in carbontetrachloride. After stirring approximately for 4 hours, the mixture canbe filtered through a pad of silica gel few times, and concentrate togive the title compound.

Step G: Preparation of 4-(3-(2,6-Dimethylbenzyloxy)phenyl)-3-butenoicacid

To a solution of Ethyl 4-(3-(2,6-dimethylbenzyloxy)phenyl)-3-butenoate(Step F) in abs ethanol at low temp, add aqueous sodium hydroxide, afteran hour, concentrate and purify by flash chromatography on silica gelcolumn (chloroform:methanol 95:5 spiked with acetic acid).

BIOLOGICAL ACTIVITY EXAMPLES Example A Compound AH Improves MetabolicAbnormalities in Insulin-Dependent Diabetes

Streptozotocin (STZ) is a toxin that selectively destroysinsulin-producing pancreatic beta cells, and is widely used to induceinsulin-dependent diabetes in experimental animals.

Female Balb/C mice (8 weeks old; 18-20 grams body weight) were treatedwith streptozotocin (STZ) (50 mg/kg i.p. on each of five consecutivedays). Fourteen days after the last dose of STZ, blood glucose wasmeasured to verify that the animals were diabetic, and the mice weredivided into two groups of 5 animals each, one group receiving CompoundAH (250 mg/kg) daily by oral gavage, and the other receiving vehicle(0.75% hydroxypropylmethylcellulose, a suspending agent, in water). Agroup of nondiabetic mice from the same cohort that did not receive STZwere also monitored. Blood samples were taken periodically fordetermination of blood glucose concentrations, and body weights werealso recorded.

After several weeks of treatment, blood glucose concentrations in micetreated with oral Compound AH began to decrease toward baseline, whereasblood glucose in the vehicle-treated control animals continued to rise.Body weights and blood glucose, triglyceride and cholesterolconcentrations 14 weeks after the beginning of drug treatment are shownin Table 1.

TABLE 1 Serum chemistries and body weights in streptozotocin diabeticmice treated with oral Compound AH for 14 weeks Glucose TriglyceridesCholesterol Body Weight Group mg/dL mg/dL mg/dL (g) Nondiabetic + 138 ±6  88 ± 9 88 ± 6   21 ± 0.6 Vehicle Diabetic + 615 ± 46 154 ± 16 133 ±6  17.5 ± 1.0  Vehicle Diabetic + 207 ± 12  62 ± 7* 82 ± 2* 21.7 ± 0.8*Compound AH *= significantly different from STZ Diabetic group, P < .001

Oral Compound AH treatment resulted in significant amelioration ofmetabolic abnormalities associated with insulin-dependent diabetes.

Example B Oral Compound AH Improves Survival of Mice with LethalInsulin-Dependent Diabetes

Female Balb/C mice (14 weeks old) were treated with a single dose ofstreptozotocin (175 mg/kg i.p.) to induce severe insulin-dependentdiabetes. Seven days later, mice were divided into three treatmentgroups: Compound AH, pioglitazone, and vehicle. Mice were treated dailyvia oral gavage, and survival was monitored over time.

TABLE 2 Survival at 12 weeks Groups Survivors Vehicle 0/5 Pioglitazone30 mg/kg/day 2/5 Compound AH 250 mg/kg/day 4/5

All of the diabetic animals treated with oral vehicle died of severe,uncontrolled diabetes. Two of five animals treated with pioglitazone, anantidiabetic insulin sensitizer used to treat humans withnoninsulin-dependent diabetes, were alive at 12 weeks, but had lost15-20% of their body weight. Four of the five animals treated with oralCompound AH were alive at 12 weeks, and their body weights recovered andwere maintained in the normal range.

Example C Oral Compound AA Reduces Mortality in Severe Insulin-DependentDiabetes

Female balb/C mice (19 wks of age at start of experiment) werechallenged with multiple high doses of STZ (75 mg/kg i.p. on 5consecutive days). Animals were then divided in two groups (20mice/group) matched for severity of diabetes. Four days after the lastdose of STZ, treatments were initiated. One group received Vehicle (0.4ml of 0.75% HPMC, p.o.), and the other group received oral COMPOUND AA(30 mg/kg/day). After three weeks of daily treatment, cumulativemortality in the Vehicle Control group was 19/20 mice. In contrast, only5/20 of the COMPOUND AA mice died during this time.

Example D Compound AH Reduces the Incidence of Spontaneous Diabetes andMortality in NOD Mice

A substantial proportion of NOD (“non-obese diabetic”) mice developinsulin-dependent diabetes as a consequence of spontaneous autoimmunedestruction of pancreatic islet cells. Two groups of 20 NOD mice (6weeks old) were treated daily with either oral Vehicle (0.4 ml of 0.75%hydroxypropyl methylcellulose in water; HPMC) or Compound AH (200mg/kg/day) suspended in HPMC. The incidence of mortality due tospontaneous development of severe insulin-dependent diabetes wasmonitored over a period of seven months. At the end of this time, 13/20mice treated with vehicle had died of uncontrolled diabetes, whereasonly 5/20 mice treated with Compound AH had died.

Example E Compound AW Reduces Hyperglycemia and Hyperlipidemia, andAmeliorates Fatty Liver Disease in ob/ob Obese Diabetic Mice

Ob/ob mice have a defect in the gene for leptin, a protein involved inappetite regulation and energy metabolism, and are hyperphagic, obese,and insulin resistant. They develop hyperglycemia and fatty liver.

Male lean (ob/+heterozygote) and obese (ob/ob homozygote) C57BL/6 miceapproximately 8 weeks of age were obtained from Jackson Labs (BarHarbor, Me.) and randomly assigned into groups of 5 animals such thatbody weights and blood glucose concentrations were similar betweengroups. All animals were maintained under the control of temperature (23C), relative humidity (50±5%) and light (7:00-19:00), and allowed freeaccess to water and laboratory chow (Formulab Diet 5008, Quality LabProducts, Elkridge, Md.). Blood glucose was routinely determined withglucose test strips and a Glucometer Elite XL device (BayerCorporation). At selected time points, blood samples (˜100 microliters)were obtained with a heparinized capillary tube via the retro-orbitalsinus for serum chemistry analysis. Serum chemistry (glucose,triglycerides, cholesterol, BUN, creatinine, AST, ALT, SDH, CPK and freefatty acids) analyses were performed on a Hitachi 717 Analyzer, andplasma insulin and pancreatic insulin were measured by anelectrochemiluminescent immunoassay (Origen Analyzer, Igen, Inc.,Gaithersburg, Md.).

Groups of ob/ob mice were divided into treatment cohorts as indicatedbelow, and given daily oral doses of Compound AW (10, 30, 100, 150 or300 mg), rosiglitazone (1, 3, 10 or 30 mg), or pioglitazone (30 or 100mg). The latter two compounds are insulin-sensitizing drugs used in thetreatment of human patients with non-insulin dependent diabetesmellitus, and are used as comparators for efficacy and safety ofcompounds of the invention. The dose ranges of compounds in thisexperiment were chosen to include both suboptimal and potentiallysupraoptimal doses.

Compound AW produced reduction in blood glucose comparable to thatachieved with pioglitazone and rosiglitazone, as shown in Table 3. Atdoses of 100 to 300 mg/kg/day, Compound AW reduced serum triglyceridesand fatty acids better than did either rosiglitazone or pioglitazone attheir optimum antihyperglycemic doses.

TABLE 3 Effect of Compound AW, pioglitazone (PG) and rosiglitazone (RSG)on serum glucose, triglycerides, and free fatty acids in ob/ob mice FreeFatty Glucose ± SEM Triglycerides ± SEM Acids ± SEM Group mg/dL mg/dLmicromoles/L ob/+ 268.6 ± 12.9 111.6 ± 12.0   2216 ± 197.4 ob/ob 384.2 ±53.8  106.6 ± 2.909   3399 ± 345.6 AW-10 369.6 ± 62.5 115.6 ± 7.8 3697.4 ± 357.8 AW-30 280.2 ± 46.7 96.4 ± 7.3 2552.2 ± 334.7 AW-100   286± 47.1 66.2 ± 5.9  1476 ± 82.1 AW-150 188.6 ± 28.8 72.6 ± 5.6   1481 ±158.8 AW-300 128.4 ± 8.8  63.6 ± 3.4 1452.6 ± 111.1 PG-30 188.2 ± 21.4111.2 ± 7.5    2606 ± 139.2 PG-100 174.6 ± 11.5 95.2 ± 4.8 1983.4 ±66.1  RSG-1 142.75 ± 8.8  109.75 ± 4.4  2090.75 ± 67.7  RSG-3 190.2 ±12.7 107.8 ± 3.8  2317.6 ± 85.3  RSG-10 188.2 ± 21.4 111.2 ± 7.5  2606.4± 139.2 RSG-30 174.6 ± 11.5 95.2 ± 4.8 1983.4 ± 66.1 

Ob/ob mice develop chronic inflammatory fatty liver disease and areconsidered to be an animal model for nonalcoholic steatohepatitis(NASH), a condition which can lead toward progressive cirrhosis andliver dysfunction. In NASH, fat accumulation increases thesusceptibility of the liver to inflammatory injury. One characteristicsign of NASH in patients is, in the absence of viral infection oralcoholism, elevated levels in serum of enzymes that are released fromdamaged hepatocytes, e.g. alanine aminotransferase (ALT), aspartateaminotransferase (AST), and sorbitol dehydrogenase (SDH). These enzymesare elevated in ob/ob mice as a consequence of fatty liver and secondaryinflammation. In Table 4, ALT, AST, and SDH in serum samples from micetreated with Compound AW, pioglitazone, and rosiglitazone are shown, asare enzyme levels in serum from normal lean mice and from diabeticcontrol mice treated only with vehicle. ALT, AST and SDH aresignificantly elevated in obese diabetic ob/ob mice compared to leanmice. Compound AW treatment at doses ranging from 30 mg/kg/day to 300mg/kg/day resulted in a dose-dependent decrease in serum liver enzymes.In contrast, pioglitazone (30 and 100 mg/kg/day) and rosiglitazone (1 to30 mg/kg/day) induced an elevation in ALT and AST and did not changeSDH. The serum liver enzyme profiles correlated with liver histology.Vehicle-treated ob/ob obese diabetic mice had marked fat accumulation inthe liver in discrete intracellular droplets. Daily Compound AWtreatment for 4 weeks caused a marked reduction in liver fat droplets,whereas neither pioglitazone nor rosiglitazone reduced the size ordensity of fat droplets in the hepatocytes.

TABLE 4 Effect of Compound AW, pioglitazone and rosiglitazone on serumenzyme indicators of liver injury SDH (U/L) ± Group ALT (U/L) ± SEM AST(U/L) ± SEM SEM Lean 106.4 ± 16.3 25.6 ± 2.7 23.2 ± 4.5 Diabetic 447.2 ±63.4  645.6 ± 104.8  745.8 ± 102.4 2022-10 483.8 ± 81.9  653.4 ± 104.8626.8 ± 93.8 AW-30 320.2 ± 46.2 399.6 ± 74.4 333.0 ± 66.9 AW-100 202.8 ±38.0 143.8 ± 30.4 121.2 ± 14.1 AW-150 149.2 ± 15.6 185.8 ± 26.0 166.2 ±20.0 AW-300 188.2 ± 10.3 335.4 ± 44.8 207.0 ± 29.3 PG-30 713.6 ± 80.6 1024 ± 88.7 782.0 ± 70.6 PG-100 646.0 ± 56.1 901.0 ± 49.3 603.0 ± 27.3RSG-1 668.8 ± 42.9 798.0 ± 73.8 644.5 ± 51.6 RSG-3 716.6 ± 56.6 853.8 ±43.8 615.4 ± 38.6 RSG-10 713.6 ± 80.5 1024.0 ± 88.7  782.0 ± 70.6 RSG-30646.0 ± 56.1 901.2 ± 49.3 603.0 ± 27.3

The ob/ob Mice gained body weight during the four week treatment period.As is shown in Table 5, pioglitazone and rosiglitazone exacerbatedweight gain relative to vehicle-treated mice, whereas Compound AWinduced a dose-dependent attenuation of weight gain.

TABLE 5 Effect of Compound AW, Pioglitazone and Rosiglitazone on bodyweight gain of ob/ob mice Groups Mean body weight gain (grams) HPMC(Vehicle) +7.4 AW - 3 mg/kg/day +7.3 AW - 10 mg/kg/day +6.7 AW - 30mg/kg/day +6.4 AW - 100 mg/kg/day +3.4 AW - 150 mg/kg/day +4.6 AW - 300mg/kg/day −0.7 PG - 30 mg/kg/day +10.0 PG - 100 mg/kg/day +13.6 RSG - 1mg/kg/day +8.2 RSG - 3 mg/kg/day +8.5 RSG - 10 mg/kg/day +11.0 RSG - 30mg/kg/day +12.0

Example F Acute Hypoglycemic Effects of Compounds of the Invention inDiabetic Mice: Experiment 1

Compounds of the invention display acute antihyperglycemic activity inanimals with non insulin-dependent diabetes.

Male ob/ob diabetic mice were randomized into groups of five animalseach. Body weights were 50-55 g and blood glucose was approximately 300mg/dL in the fed state. A single oral dose of a test substance suspendedin 0.5% carboxymethylcellulose vehicle was administered by gavage. Bloodglucose was measured in blood droplets obtained by nicking a tail veinwith a razor using glucometer test strips and a Glucometer Elite XLdevice (Bayer) at 0, 0.5, 2, 4, 6 and 18 hours after the initial dosing.A 10% reduction in blood glucose versus oral vehicle is considered apositive screening result. Blood glucose reductions were generallymaximal at 6 hours after drug administration. Table 6: Acutehypoglycemic effect of compounds of the invention in ob/ob obesediabetic mice.

Blood Glucose % Reduction vs Treatment Group After 6 hours ControlVehicle 297 ± 35   0.0 ± 11.8 Compound AA 242 ± 25 −18.5 ± 8.4 CompoundAB 181 ± 19 −39.1 ± 6.4 Compound AF 314 ± 32  −24.6 ± 7.7* Compound AG222 ± 23 −25.3 ± 7.7 Compound AH 223 ± 11 −24.9 ± 3.7 Compound AI 255 ±9  −14.1 ± 3.0 Compound AJ 190 ± 14 −36.0 ± 4.7 Compound AK 210 ± 10−29.3 ± 3.4 Compound AL 168 ± 13 −43.4 ± 4.4 *Initial blood glucose inthis group was 416 ± 29 mg/dL and the 6 hour reading is normalized tothat initial value. In all other groups in this experiment, mean initialblood glucose was ≦300 mg/dL.

Example G Acute Hypoglycemic Effects of Compounds of the Invention inDiabetic Mice: Expt 2

Compounds of the invention display acute antihyperglycemic activity inanimals with noninsulin-dependent diabetes.

Male ob/ob mice (50-55 grams; blood glucose ˜300 mg/dL) were dividedinto groups of five animals each, and given a single oral dose of testdrug (250 mg/kg) suspended in 0.5% carboxymethylcellulose vehicle; acontrol group received oral vehicle alone.

Six hours after oral administration of test drugs or vehicle (control),blood samples were obtained from a tail vein and glucose content wasdetermined with a glucometer.

TABLE 7 Acute hypoglycemic effect of compounds of the invention in ob/obobese diabetic mice Blood Glucose % Reduction vs Treatment Group after 6hours Control Vehicle Control 305 ± 20  0.0 ± 5.0 mg/dL Compound AN 152± 11 −50.2 ± 4.5% Compound AQ 220 ± 17 −27.9 ± 4.2% Compound AR 179 ± 14−41.3 ± 4.2% Compound AS 167 ± 28 −45.2 ± 2.0% Compound AT 198 ± 28−35.1 ± 2.3% Compound AU 224 ± 26 −26.6 ± 2.8% Compound AV 207 ± 23−32.1 ± 3.0% Compound AW 143 ± 15 −53.1 ± 3.1% Compound AX 165 ± 23−45.9 ± 2.4% Compound AY 185 ± 21 −39.3 ± 2.9% Compound AZ 186 ± 10−39.0 ± 6.1%

Oral treatment with compounds of the invention elicits an acuteantihyperglycemic effect in obese diabetic mice.

Example H Antidiabetic Effects of Compounds of the Invention in db/dbMice

Db/db mice have a defect in leptin signaling, leading to hyperphagia,obesity and diabetes. Moreover, unlike ob/ob mice which have relativelyrobust islets, their insulin-producing pancreatic islet cells undergofailure during chronic hyperglycemia, so that they transition fromhyperinsulinemia (associated with peripheral insulin resistance) tohypoinsulinemic diabetes.

Male db/db mice were given daily oral treatments with vehicle (0.75%hydroxypropylmethylcellulose) or antidiabetic compounds as indicatedbelow. Blood samples were obtained via the retro-orbital sinus for serumchemistry analysis, or via the tail vein for glucose measurement with atest strip and glucometer.

After four weeks of daily oral dosing, Compound AW and Compound BHelicited a significant reduction in blood glucose. While pioglitazonedid initially reduce blood glucose over the first 3 weeks, its activityhad largely failed at the 4 week time point and thereafter. The dose ofpioglitazone used in this experiment was reported in the literature tobe a maximally-effective dose for treatment of db/db mice (Shimaya etal. (2000), Metabolism 49:411-7).

TABLE 8 Groups Glucose mg/dL Glucose (% of Control) Vehicle (Control)562 ± 24  100 ± 4  Compound AW - 150 mg/kg 313 ± 34* 56 ± 6* CompoundBH - 150 mg/kg 229 ± 49* 41 ± 9* Pioglitazone - 100 mg/kg 558 ± 28  99 ±5  *Less than Vehicle Control value, p < .05

In a second experiment in db/db mice, antidiabetic activity of CompoundBI was compared with that of rosiglitazone. After 8 weeks of treatment,blood glucose and triglycerides were significantly lower in animalstreated with either Compound BI or rosiglitazone, compared tovehicle-treated controls. The rosiglitazone dose used in this study wasreported in published literature as the optimum dose for late stagedb/db mice (Lenhard et al., (1999) Diabetologia 42:545-54). Groupsconsisted of 6-8 mice each.

TABLE 9 Groups Glucose (mg/dL) Triglycerides (mg/dL) Vehicle (Control)686 ± 47  147 ± 13  Rosiglitazone - 20 mg/kg 343 ± 38*  89 ± 16*Compound BI - 150 mg/kg 254 ± 30* 99 ± 8* *= Less than Vehicle Controlvalue, P < .05 (One-way ANOVA)

Example I Antidiabetic Effects of Compounds of the Invention in db/dbMice

db/db mice have a defect in leptin signaling, leading to hyperphagia,obesity and diabetes. Moreover, unlike ob/ob mice on a C57BL/6Jbackground, db/db mice on a C57BL/KS background undergo failure of theirinsulin-producing pancreatic islet β cells, resulting in progressionfrom hyperinsulinemia (associated with peripheral insulin resistance) tohypoinsulinemic diabetes.

Male obese (db/db homozygote) C57BL/Ksola mice approximately 8 weeks ofage, were obtained from Jackson Labs (Bar Harbor, Me.) and randomlyassigned into groups of 5-7 animals such that the body weights (50-55 g)and serum glucose levels (≧300 mg/dl in fed state) were similar betweengroups; male lean (db/+heterozygote) mice served as cohort controls. Aminimum of 7 days was allowed for adaptation after arrival. All animalswere maintained under controlled temperature (23° C.), relative humidity(50±5%) and light (7:00-19:00), and allowed free access to standard chow(Formulab Diet 5008, Quality Lab Products, Elkridge, Md.) and water.

Treatment cohorts were given daily oral doses of (1%hydroxypropylmethylcellulose), Compounds BI, BO, BP, BQ or BR for 2weeks. At the end of the treatment period 100 μl of venous blood waswithdrawn in a heparinized capillary tube from the retro-orbital sinusof db/db mice for serum chemistry analysis.

Effects of compounds of the invention on nonfasting blood glucose areshown in Table 10; effects on serum triglycerides and free fatty acidsare shown in Table 11.

TABLE 10 The effects of Compounds BI, BO, BP, BQ or BR on blood glucosein the db/db mouse model Glucose Groups Glucose mg/dL (% of Control)Vehicle (Control) 632 ± 19 100 ± 3  BI - 150 mg/kg  279 ± 35* 44 ± 6*BI - 100 mg/kg  423 ± 53* 67 ± 8* BO - 100 mg/kg 586 ± 58 93 ± 9  BP -100 mg/kg 629 ± 86 99 ± 14 BQ - 100 mg/kg  473 ± 49* 75 ± 7* BR - 82mg/kg 703 ± 64 111 ± 10  Blood glucose levels in lean, nondiabetic db/+heterozygote mice were 225 ± 15 mg/dL

TABLE 11 Effect of Compounds BI, BO, BP, BQ or BR on serum glucose,triglycerides, and free fatty acids in db/db mice Triglycerides ± SEMFree Fatty Acids ± SEM Group (mg/dL) (μM) Lean 142.4 ± 6.3  2577.6 ±80.8  Diabetic 444.3 ± 57.3 4044.9 ± 158.5 BI-150 103.6 ± 8.3  2234.0 ±162.6 BI-100 134.0 ± 13.1 2999.9 ± 98.7  BO-100 261.1 ± 24.3 3766.3 ±234.5 BP-100 302.1 ± 28.1 3772.6 ± 182.5 BQ-100 131.6 ± 20.7 2825.9 ±110.9 BR-82 253.0 ± 32.0 3653.4 ± 207.5

Example J Antidiabetic Effects of Compounds of the Invention in db/dbMice

db/db mice have a defect in leptin signaling, leading to hyperphagia,obesity and diabetes. Moreover, unlike ob/ob mice on a C57BL/6Jbackground, db/db mice on a C57BL/KS background undergo failure of theirinsulin-producing pancreatic islet cells, resulting in progression fromhyperinsulinemia (associated with peripheral insulin resistance) tohypoinsulinemic diabetes.

Male obese (db/db homozygote) C57BL/Ksola mice approximately 8 weeks ofage, were obtained from Jackson Labs (Bar Harbor, Me.) and randomlyassigned into groups of 5-7 animals such that the body weights (50-55 g)and serum glucose levels (≧300 mg/dl in fed state) were similar betweengroups; male lean (db/+heterozygote) mice served as cohort controls. Aminimum of 7 days was allowed for adaptation after arrival. All animalswere maintained under controlled temperature (23° C.), relative humidity(50±5%) and light (7:00-19:00), and allowed free access to standard chow(Formulab Diet 5008, Quality Lab Products, Elkridge, Md.) and water.

Treatment cohorts were given daily oral doses of Vehicle (1%hydroxypropylmethylcellulose), Compounds BI, BS, BT, BU, BV orFenofibrate for 2 weeks. At the end of the treatment period 100 μl ofvenous blood was withdrawn in a heparinized capillary tube from theretro-orbital sinus of db/db mice for serum chemistry analysis.

Effects of compounds of the invention on nonfasting blood glucose areshown in Table 12; effects on serum triglycerides and free fatty acidsare shown in Table 13.

TABLE 12 The effects of compounds BI, BS, BT, BU, BV and fenofibrate indb/db mice Glucose Groups Glucose mg/dL (% of Control) Vehicle (Control)692.5 ± 55.4 100 ± 8  BI - 100 mg/kg  347.0 ± 43.1*  50 ± 6* BS - 93mg/kg  372.0 ± 53.8*  54 ± 8* BT - 107 mg/kg 684.3 ± 63.6 99 ± 9 BU -128 mg/kg 533.3 ± 46.7 77 ± 7 BV - 115 mg/kg 789.5 ± 38.9 114 ± 6 Fenofibrate - 113 mg/kg 563.2 ± 49.0 81 ± 7 Blood glucose levels inlean, nondiabetic db/+ heterozygote mice were 208.5 ± 6.6 mg/dL

TABLE 13 Effect of compounds BI, BS, BT, BU, BV and Fenofibrate on serumtriglycerides and free fatty acids in db/db mice Triglycerides ± SEMFree Fatty Acids ± SEM Group (mg/dL (μM) Lean 114.2 ± 8.7  2315.8 ±238.3 Vehicle 232.8 ± 20.7 3511.8 ± 257.6 BI 77.8 ± 5.3 1997.2 ± 196.4BS 132.0 ± 15.2 2867.4 ± 267.7 BT 211.5 ± 21.5 3897.7 ± 291.3 BU 172.5 ±9.9  3587.0 ± 156.3 BV 153.2 ± 14.2 3373.8 ± 233.6 Fenofibrate 109.3 ±9.1  3318.5 ± 208.7

Example K Attenuation of Cataractogenesis of Compounds of the Inventionin Zucker Diabetic Fatty (ZDF) Rats

Cataracts are one of the leading causes of progressive vision declineand blindness associated with ageing and diabetes, and the Zuckerdiabetic fatty (ZDF) model has many similarities with humancataractogenesis, including biochemical changes and oxidative stress inthe lens. These rats, however, undergo cataractogenesis typicallybetween 14-16 weeks of age.

Male ZDF rats and their aged-match Zucker lean (ZL) counterparts (fa/+or +/+) were obtained from Genetic Models, Inc. (Indianapolis, Ind.)aged 12 weeks and acclimatized for 1 week prior to study. All animalswere maintained under controlled temperature (23° C.), relative humidity(50±5%) and light (7:00-19:00), and allowed free access to standard chow(Formulab Diet 5008, Quality Lab Products, Elkridge, Md.) and tap waterad libitum. Treatment cohorts were given a daily oral dose of vehicleand 100 mg/kg of BI or BH for 10 weeks. Body weights and blood glucosewere routinely determined (once a week, usually around 10:00 A.M.) fromtail bleeds with glucose test strips and a Glucometer Elite XL device(Bayer Corporation). At the end of the treatment period 100 μl of venousblood was collected (usually 10:00 A.M.) in a heparinized tube from thetail vein for serum chemistry analysis (Anilytics, Inc., Gaithersburg,Md.). Serum chemistry (glucose (GL), triglycerides (TG), aspartateaminotransferase (AST), alanine aminotransferase (ALT), sorbitoldehydrogenase (SDH), and free fatty acids (FFA)) analyses were performedon a Hitachi 717 Analyzer (Anilytics, Inc., Gaithersburg, Md.). Plasmainsulin was measured by an electrochemiluminescent immunoassay, ECL(Origen Analyzer, Igen, Inc., Gaithersburg, Md.). The animals weresacrificed and tissues and/or organs (lens and liver) were extirpated,weighed (wet weight) and processed for biochemical analyses.Malondialdehyde (MDA), a major product of lipid peroxidation was assayedin lenses according to Ohkawa et al (1979), Analytical Biochem 95,351-358).

Table 14 shows the incidence of visible cataracts in the eyes of the ZDFrats. Table 15 indicates additional quantitative indices ofcataractogenesis in the same animals.

TABLE 14 Attenuation of cataractogenesis by Compounds BH and BI in ZDFrats. Cataract Formation % Protection Animal Groups N Left Eye Right EyeLeft Eye Right Eye Vehicle-Control 6 6/6 6/6 0 0 BI 6 3/6 1/6 50 83 BH 64/6 5/6 33 17 Lean 4 0/4 0/4 N/A N/A

TABLE 15 Attenuation of cataractogenesis by BH and BI in ZDF rats.Lenticular Size (mm) MDA Weight (mg) Right nmol/g Groups Left Lens RightLens Left Lens Lens lens LEAN 51.2 ± 3.5 59.0 ± 0.4 3.8 ± 0.2 3.9 ± 0.10.4 ± 0.0 Vehicle 15.1 ± 1.4 16.8 ± 1.7 1.9 ± 0.1 2.0 ± 0.2 2.4 ± 0.2 BI38.1 ± 7.3** 54.9 ± 1.2* 3.4 ± 0.2* 3.8 ± 0.1* 0.8 ± 0.1‡ BH 27.0 ± 7.220.0 ± 6.6 2.5 ± 0.3 2.1 ± 0.4 1.9 ± 0.2 Data are means ± SEM. *p < 0.05compared with the vehicle-controls (diabetic) and Compound BH-treatedgroups, respectively; **p < 0.05 compared with vehicle-controls; ‡p <0.05 compared with vehicle-controls and Compound BH right lenses,respectively (One Way ANOVA, Tukey Test) All pairwise MultipleComparison.

Example L Oral BI and BL Lower Circulating Triglycerides, Free FattyAcids, Insulin and Leptin in High Fat-Fed C57Bl/6J Mice

The high fat-fed mouse is a model for the hypertriglyceridemia and highcirculating fatty acid levels, and the insulin and leptin resistancethat are found in people at risk for and with obesity, diabetes,cardiovascular disease and other disorders. Male C57Bl/6J mice,approximately 8 weeks of age, were randomly assigned into groups of 6animals. They were maintained under controlled temperature (23° C.),relative humidity (50±5%) and light (7:00-19:00), and allowed freeaccess to food and water ad libitum. Mice were fed a high-fat diet (dietnumber D12451, containing 45% of calories as fat (Research Diets, NewBrunswick, N.J.)) for 6 weeks. After the 6 weeks, groups of micereceived either vehicle (hydroxymethylcellulose), BI, BL, Wy14,643 orrosiglitazone by oral gavage at the indicated doses for an additional 4weeks while continuing on the high-fat diet. Plasma chemistries(Anilytics, Inc., Gaithersburg, Md.) were assayed after 2 weeks of drugtreatments. Plasma serum insulin (FIG. 1) and leptin (FIG. 2) weremeasured by an electrochemiluminescent immunoassay (Origen Analyzer,Igen, Inc., Gaithersburg, Md.) after 4 weeks of drug treatments.

BI and BL were effective at lowering serum triglycerides and free fattyacids as well as insulin and leptin serum levels. Serum values from micefrom the same cohort (“lean controls”) that were maintained on regularlab chow (Formulab Diet 5008, Quality Lab Products, Elkridge, Md.) areshown for comparison.

TABLE 16 Triglycerides Free Fatty (mg/dL) Acids (umol/L) Vehicle  135 ±40.1   1686 ± 359.3 BI (10 mg/kg) 68.8 ± 5.7    1227 ± 193.7 BI (30mg/kg) 66.5 ± 14.7   1292 ± 231.4 BI (100 mg/kg) 37.4 ± 8.3   992.8 ±172.1 BL (10 mg/kg)   80 ± 12.2 1571.8 ± 100.9 BL (30 mg/kg) 66.4 ± 13.71413.2 ± 228.7 BL (100 mg/kg)  41 ± 5.6 1133.5 ± 132.7 Rosiglitazone (1mg/kg) 76.6 ± 16.5   1537 ± 256.3 Rosiglitazone (3 mg/kg) 103.2 ± 10.8 1833.2 ± 169.8 Rosiglitazone (10 mg/kg) 129.5 ± 48.7  1810.3 ± 595  Rosiglitazone (100 mg/kg)  88 ± 7.2 1568.5 ± 197   Wy14643 (10 mg/kg)70.6 ± 10.8 1512.2 ± 172.9 Wy14643 (30 mg/kg)   88 ± 12.5 1676 ± 237Wy14643 (100 mg/kg) 88.4 ± 18.8 1839.8 ± 154.8 Rosi (3 mg/kg) + Wy1464354.3 ± 10.5 1649.7 ± 260.5 (100 mg/kg)

Example M Oral BI lowers circulating triglycerides, free fatty acids,insulin and leptin in high fat-fed Sprague Dawley rats

The high fat-fed rat is a model for insulin and leptin resistance.Sprague-Dawley rats have an intact leptin system and respond to a highfat diet with hyperinsulinemia due to a downregulation of the normalinsulin response in peripheral tissues such as liver, adipose tissue andmuscle

Male Sprague-Dawley rats, approximately 17 weeks of age, were obtainedfrom Jackson Labs (Bar Harbor, Me.) and randomly assigned into groups of5-7 animals; the body weights were similar between groups. All animalswere maintained in a temperature-controlled (25° C.) facility with astrict 12 h light/dark cycle and were given free access to water andfood. Rats were fed a high-fat diet (diet number D 12451 (containing 45%of calories as fat), Research Diets, New Brunswick, N.J.) for one monthprior to drug treatment.

Groups of 6 Sprague-Dawley rats were treated with a single daily dose ofvehicle (hydroxymethylcellulose), BI (10, 30 and 100 mg/kg), orrosiglitazone (3 mg/kg) for 6 weeks while maintaining the high-fat diet.At the indicated time points, blood samples (˜100 μl) were obtained viathe tail vein for serum chemistry analysis.

BI (30 mg/kg) reduced serum insulin, triglycerides; BI at all dosesreduced free fatty acids.

TABLE 17 Effect of BI and rosiglitazone on serum glucose, insulin,triglycerides and free fatty acids in high-fat fed Sprague-Dawley ratsGlucose Insulin Triglycerides Free Fatty Acids Group (mg/dL) (ng/ml)(mg/dL) (μMol/L) Lean 123.8 ± 7.0 0.72 ± 0.1 179.0 ± 72.3 743.5 ± 57.4Vehicle 122.3 ± 5.9 1.78 ± 0.3 200.7 ± 39.2  942.5 ± 181.0 BI-10 117.3 ±8.8 2.18 ± 0.9 183.7 ± 58.4  923.7 ± 161.3 BI-30  127.3 ± 22.2 1.46 ±0.2 129.3 ± 20.0 738.7 ± 50.0 BI-100  19.3 ± 3.5 1.79 ± 0.2 171.7 ± 33.1725.7 ± 87.5 RG-3 119.8 ± 5.4 1.57 ± 0.2 134.2 ± 15.2 758.8 ± 61.0

The invention claimed is:
 1. A method for treating a mammalian subjectwith a condition selected from the group consisting of insulinresistance syndrome and Type II Diabetes, comprising administering tothe subject an amount of4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyric acid or apharmaceutically acceptable salt thereof effective to treat thecondition.
 2. The method of claim 1, wherein the4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyric acid orpharmaceutically acceptable salt thereof is administered orally.
 3. Themethod of claim 1, wherein the subject is a human.
 4. The method ofclaim 3, wherein the 4-(3-(2,6-dimethylbenzyloxy)phenyl)-4-oxobutyricacid or pharmaceutically acceptable salt thereof is administered in anamount from one milligram to four hundred milligrams per day.